Hes-1 的類小泛素化修飾可調節 Hes-1 蛋白質的穩定及 GluR1 的表現 - 政大學術集成
95
0
0
全文
(2) /(Hes-1 %13 GluR1 #. and GluR1 expression. 學 ‧. ‧ 國. 政 治 大 Sumoylation of Hes-1 立regulates the protein stability of Hes-1. er. io. sit. y. Nat. n. al v ƪĎʉŦŨΑ ni Ch. U. engchi ƏϋɔɑĄ5ʬʦ2 ͼšʜʦ2. ;˜Ⱥ101 À7 `26 _.
(3) ͶЌ ĦÀşƪĎņɦ ʯʎΔòƱş͆+Л ʢƺ̇ЌƏϋɔɑ¸ʎƙǬ ˱:γƓƟ+ńƓƪĎ+˖=øşƏϋPɖˋ ʷĢ˖ø NJƆºşƪĎϿ ͋ ѭø|o± ̡ϴčşϊŭĹljϊʁ ʢƈ ƺ̇Ќ1̯ĸǗȥ͎γ]ş Ȑ˖=øы˦̆đ ġγ]ȓĂò˗ ¸ʎĦÀş̼ˎ; ̇Ќ͌ѪƁϊŭĻlj¸͌Ѫ+˖=øʢ3ş̓϶ Į çøʮü^± ЌЌіΞϊĻ
(4) ̙Ώϊŭ
(5) ˳ϊŭ
(6) 3ǎϊŭ
(7) ɼ¾ϊŭ
(8) ƅêϊŭ
(9) ļÚϊŭ
(10) ƅËϊŭ
(11) Řʡϊŭ NJćѮ NJćѮ:͇Ĩ̗şƲ ZƏϋ Ěj¸Ĉ Çø͎α̎P͌Ѫ+ɰ͞ʳşǬNj Įçø Ț̭ċȸ е ¯Ǭ -ƺ̇Ќȩǔ¸ȵŝ͌Ѫ+şĮç ȧ ͌Ѫ+Pα̎+ -Ų ɃЌЌϊŭĻlj¸ʎƙʽ˱͎øƟ+ş̘ѡ ѭøş͡2ɦ¸ѣΙ;ƃ ̼ ČÆ͌ѪƁϊŭĻljoPÍǠşƲZƏϋ ʯČÆʎΥγ]şί È. 立. 政 治 大. ‧. ‧ 國. 學. -ƺ̇Ќȇ̞ƫϊƪĎņņϦş.Ĺ Įçøʃɮʉ¹]Ǯ+şȸе ʢƈ̇ЌøşǛ"͎øşdzȾP¨Z¨̆ş\Ǝ˝ø ѭøȓʾ˝Ζź ˬ ¯Ǭ-ƺ̇ЌøĖǫşŎOljoPтøşϊŭĻlj˖=øʢ3ş\Ǝͯ ̓϶ ¸øĈ Çǂ͎ǨýPϷ&şǬNj ʚ˝øβϑ̢ćƿ̋vˋ ʢƈ¸ È̇Ќ͡2ɦʎƙǬ˱ņÆϸç̼øPчZøş" ЫoÈγ]э˖ßlj. n. er. io. sit. y. Nat. al. Ch. engchi. I. i Un. v.
(12) ;]͓ƺ Э ђ ƈ Ǎ ͂ Þ (post-translational modifications). yíȼL. (methylation)
(13) ЃL(phosphorylation)
(14) ŚȋL(ubiquitination)
(15) щ5Śȋ LǍ͂(sumoylation) ˑ̼{ÆƪĎƏvщ5ŚȋLǍ͂|oβ̝Ζʅ δşмļƃ ˬÏβ̝ʉ¹ɽƵFQϹ ģ»ɽƵǯ̸ϥÞ
(16) DNA ή ͵
(17) β̝ЭϭÞ
(18) ƖÒѫIб
(19) ȘǦ˹;
(20) ɽƵ˫ʽβɌ
(21) DNA Ǎ̪Þˑ ɯˣ щ5ŚȋLǍ͂ƓЩ ď¬şͿȋ ġщ5ŚȋʎNJʅδȓȿǍ͂ Ζʅδşlysine˂ȼ щ5ŚȋLǍ͂Ɠ NJ|ȜŃȵ͏Ǎ̼͂ˎ щ 5ŚȋLǍ͂ʐ˓ʕǤyíÆ)NJlƺşćѮ ƟL (activation) ˓³. 治 政 大 Ϳȋ˻Lş ΥƪĎlƺƓЩщ5ŚȋE3 ʐɎPIAS1 ˬÔǍ͂Þ 立 øljˋɯHairy and Enhancer of split 1 (Hes-1) ʅδ|ʇщ5ŚȋǍ͂Ʒ. (conjugation) ʐɎ (ligation) ljIåƓЩE1
(22) E2 ıE3 ʎ)͢:¯ş. ‧ 國. 學. ɂщ5ŚȋE3 ʐɎ PIAS1 ƮѧʯˈŖѭʘƀHes-1 ˬÔщ5ŚȋǍ. ‧. ͂L у͌PIAS1 şȷ͎ͯŊщ5ŚȋLǍ͂þ͖ƿƺşĒÒȧÈ< ɂщ5ŚȋΖʅδ Hes-1 ó¬+ɸ$NJ×̟ş lysine Ʈѧ ̍Āĭ. Nat. sit. y. Hes-1 ˬÔщ5ŚȋLǍ͂µÈ ʔ̼PIAS1 ņˬÔşщ5ŚȋLǍ͂|o. er. io. ġΖʅδHes-1 ʅδĂƠмļ <ƈĂˬ ćɍȗ¸ţ˱ϊʁͯȖό. n. a ; Hes-1 ˬÔщ5ŚȋLǍ͂ͯGluR1 ʅδŪɯşчŸ ͌Ѫ˓ŏѩ iv Hes-1 ǁ. l C hengchi Un ˬÔщ5ŚȋLǍ͂ġţ˱ϊʁͯȖόѧǟĚġGluR1. II. ʅδŪɯ*.
(23) Abstract There are several post-translational modifications including methylation
(24) phosphorylation
(25) ubiquitination
(26) sumoylation, etc. Previously studies indicated that sumoylation can regulate target protein stability. Sumoylation also modulates many cellular processes, including nuclear transport, DNA replication, transcription, chromosome segregation, signal transduction, cell cycle and DNA repair. Sumoylation is a process mediated by SUMOs which are attached to specific lysine residues of target proteins by the action of a series of enzymes. Sumoylation is a dynamically reversible process.. 政 治 大 which are respectively mediated by E1, E2 and E3 ligase. This study focuses 立 Sumoylation consists of three stepsactivation, conjugation and ligation,. on SUMO modification by E3 ligase. Here, we identified a new target protein,. ‧ 國. 學. Hairy and Enhancer of split 1 (Hes-1), for SUMO conjugation. The E3 ligase. ‧. deficient mutant of PIAS1 that leads to failure of Hes-1 protein sumoylation. We demonstrared that PIAS1 is involved in SUMO modification of Hes-1. In. y. Nat. Hes-1 protein on lysine 8 residue that inhibits the. sit. addition mutantion of. er. io. sumoylation of Hes-1. Therefore, sumoylation of Hes-1 regulates the protein. n. stability of Hes-1. Further astudy of the relationship i v between sumoylation of l. n. C. h e nformation Hes-1 and GluR1 in spatial memory g c h i Uindicated that spatial memory is impaired and GluR1 protein expression is decreased upon sumoylation of Hes-1.. III.
(27) ϭ ͶЌ ;]͓ƺ Ƹ]͓ƺ ϭ ͊Ç Ƹ]ЅŪ ɸ ʜ
(28) ͭγ1 ɸ ̝
(29) ϊʁͯȖό2
(30) ϊʁͯȖόşļ̠2. 政 治 大 )
(31) ȷͯϊʁͯȖόş̥ȶ3 立. !
(32) ϊʁͯȖόş͢щ2. ɸ!̝
(33) ţ˱ϊʁȖόͯǻȩȟɾФ4. ‧ 國. 學.
(34) ǻȩȟş͕ʓͯÿǝ̴Ǥ4. ‧. !
(35) ǻȩȟƨчşȵŝÔƠΘÁ5 ɸ)̝
(36) Hes-1ʅδCɼ..6. y. Nat. sit.
(37) Hes-1ȼµ˓͕ͯЭϭƟŃ6. er. io. !
(38) Hes-1ȼµŪɯşȘ̩βɌ..7. n. al )
(39) Hes-1ȼµ¸ȇ̞ˋĐďɻşŪɯ..8 iv n. C. hengchi U ɸ̝
(40) щ5ŚȋLǍ͂..9
(41) щ5Śȋ˓͕ͯIщ..9 !
(42) щ5Śȋ|ȜŃǍ͂ʕǤ10 )
(43) щ5ŚȋLǍ͂şxȓ12 ɸ?̝
(44) щ5ŚȋE3ʐɎPIAS1ʅδCɼ14
(45) PIASǛɘCɼ14 !
(46) PIAS˓͕Cɼͯщ5ŚȋLǍ͂şчŸ14 ɸG̝
(47) γ]<ƪĎşP˒ɱ16. ɸ!ʜ
(48) ͌ѪĆǪͯ^Ŗ17 ɸ ̝
(49) ɽƵȾξ(Cell culture)18
(50) ɽƵ̭Ǐ18 IV.
(51) !
(52) ɽƵȾξ18 )
(53) äǏɽƵ18 ɸ!̝
(54) ɽƵЭƖ(Transfection)19
(55) ɽƵЭƖ19 !
(56) ʅδïδɣş˛į19 ɸ)̝
(57) Ö^ρЛŖ(Western blot)19
(58) ʅδϖƃˇļPΔȣ͵19 !
(59) ɶ'!Ѹ˯ͮiѼѴτά͵ʠ20 )
(60) άѫ̿œ21
(61) ʅδ̿œЭ͘Ŗ21. 治 政 大 Co-IP)22 ɸ̝
(62) ©¯ãƦĊϕŖ (Co-immunoprecipitation, 立 ɸ?̝
(63) ͕Ƅδѫ(Plasmid construction)22. ?
(64) ãƦƖÒŖ21. ‧ 國. 學.
(65) ͮ³ʐаQϹP͕Ƅδѫ22. ‧. !
(66) Эƀ22 )
(67) 5˭δѫDNAşňį23. Nat. sit. y.
(68) Ƅ͕ļЛƮѧ<δѫ24. er. io. ?
(69) ˞ɣşŶ½24. n. ɸG̝
(70) ʅδмļIő a(Protein stability assay)24 iv. l C n U h eanimals)24 ɸ ̝
(71) ͌Ѫȵŝ(Experimental i h ngc. ɸ$̝
(72) ǻȩȟFȼµЭƖÞ (transfection).25
(73) ѫļ×[ʆͯǙͥ25 !
(74) δѫȼµͯͮ ɪĞѴɧ³ŝş͵ʠ25 )
(75) ǻȩȟCA1 ȶȻŒǝ25 ɸ ̝
(76) ȵŝÔƠ͌ѪʂdeȝƜ͌Ѫ(Morris water maze)26
(77) eȝƜ̯ѪφĆ26 !
(78) ЖЧÁeȝϊʁƜ̯Ѫ26 )
(79) Iб̥ɾФ27
(80) ̥ɾФʅδ˛į27 ?
(81) ǻȩȟɾФ<ãƦĊϕͯÖ^ρЛŖ27 V.
(82) ɸ'̝
(83) пŝş͵ʠ(Preparation of drugs)27 ɸ'̝
(84) ɻƻIő (Statistics)28 ɸ)ʜ
(85) ͌Ѫ˓ŏ29 ɸ ̝
(86) Hes-1 ʔ̼PIAS1 ˬÔщ5ŚȋLǍ͂30 ɸ!̝
(87) ɍȗщ5ŚȋLǍ͂Hes-1 şlysine ×̟33 ɸ)̝
(88) щ5ŚȋǍ͂LΎѠHes-1 ʅδмļƃ37 ɸ̝
(89) щ5ŚȋǍ͂LΎѠFŃHes-1 ʅδşмļƃ42 ɸ?̝
(90) Hes-1 ʔ̼PIAS1 ˬÔщ5ŚȋLǍ͂ΎѠHes-1 ʅδмļƃ 49 ɸG̝
(91) Hes-1 ˬÔщ5ŚȋLǍ͂β̝ţ˱ϊʁȖόPǻȩȟCA1 ȶȻ. 政 治 大 ɸʜ
(92) ȗγ60 立. GluR1 ʅδşŪɯ57 ɸ?ʜ
(93) ˓γ70. ‧ 國. 學. ȷÎ]э72. ‧. űϭ.82 űϭ
(94) Hes-1ȼµşconserved domains82. y. Nat. sit. űϭ!
(95) щ5ŚȋLǍ͂ʐ˓ʕǤ83. er. io. űϭ)
(96) PIAS1 ˓͕84. n. al űϭ
(97) Hes-1 ʅδмļƃ85 iv Ch. n engchi U. VI.
(98) ͊Ç ͊1
(99) HEK293T ɽƵFŃHes-1 ʅδ|ʔ̼PIAS1 ˬÔщ5ŚȋL Ǎ͂ 31 ͊2
(100) ɍȗHes-1 ʅδ+|ʇщ5ŚȋǍ͂şlysine ×̟ 35 ͊3
(101) ğώϖƃɜƃЭƖ͢:¯ϖƃ<Hes-1 δѫ ˇļHes-1 ʅδ ¸HEK293T ɽƵşŪɯ 38 ͊4
(102) щ5ŚȋLǍ͂ΎѠHes-1 ʅδşмļƃ 40 ͊5
(103) HEK293T ɽƵFHes-1 ʅδʔ̼PIAS1 ˬÔщ5ŚȋLǍ͎͂ ŊĨFŃHes-1 ʅδмļƃşΎѠ 44. 政 治 大 ͊7
(104) ¯ǬЭƖʘƀPIAS1 PʘƀHes-1 ńƓƮѧƀHes-1 δѫ 立. ͊6
(105) ĀĭɽƵFPIAS1 ˬÏĀĭHes-1 ʅδşмļƃ 47 ѱ͍Hes-1 ʅδşмļ 51. ‧ 國. 學. ͊8
(106) ¯ǬЭƖSUMO1 δѫPPIAS1 δѫPʘƀHes-1 ńƓƮѧƀ. ‧. Hes-1ѱ͍ʇщ5ŚȋǍ͂ƈHes-1 ʅδмļƃ 53 ͊9
(107) Hes-1 ʅδʔ̼PIAS1 ˬÔщ5ŚȋLǍ͂Ïβ̝Hes-1 ʅδ. y. Nat. sit. мļƃ 55. er. io. ͊10
(108) 3̈́ǻȩȟCA1 ȶȻFЭƖHes-1 |Āĭ3̈́şţ˱ϊʁȖ. n. al όȓ& 58 iv Ch. n engchi U. VII.
(109) Ƹ]ЅŪ AD acid domain Ala (A) alanine AMPA α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid ANOVA analysis of variance Arg (R) arginine Asn (N) asparagine APS ammonium persulfate bHLH basic helix-loop-helix BSA bovine serum albumin BMP bone morphogenic protein CA1 cornu ammonis 1. 政 治 大. CaMK Ca2+/calmodulin-dependent kinase . 立. CNS central nerve system. ‧ 國. CHX cycloheximide. 學. Cys (C) cystine. ‧. CMV cytomegalovirus. DNA deoxyribonucleic acid. Nat. sit. y. DMSO dimethyl sulfoxide. io. EGFP enhanced green fluorescent protein. al. n. FBS fetal bovine serum GluR1 glutamate receptor 1. Ch. engchi. HA hemagglutinin A Hes-1 Hairy and Enhancer of split 1 HEK293T human embryonic kidney 293T ICD intracellular domain Ile (I) isoleucine IP immunoprecipitation JNK1 c-Jun N-terminal kinase 1 kDa kilodalton µg microgram µl microliter mRNA messenger RNA VIII. er. DMEM Dulbecco's Modified Eagles Medium. i Un. v.
(110) NDSM negatively charged amino acid dependent SUMO motif NMDA N-methyl-D-aspartate NSCs neuron stem cells Lys (K) lysine PBS phosphate buffered saline PCR polymerase chain reaction PDSM phosphorylation-dependent SUMO motif PIAS1 protein inhibitor of activated signal transducer and activators of transcription 1 PINIT Pro-Ile-Asn-Ile-Thr Pro (P) proline PVDF polyvinylidene fluoride membrane RLD RING finger like zinc binding domain. 政 治 大 SAP scaffold attachment factor-A/B/acinus/PIAS 立 RNA ribonucleic acid. SD Sprague-Dawley. ‧ 國. 學. SDS sodium dodecyl sulfate SDS-PAGE SDS-PolyAcrylamide Gel Electrophoresis. ‧. Ser (S) serine. y. sit. io. siRNA small interfering RNA. n. al. er. Shh sonic hedgehog. Nat. SENP sentrin-specific proteases. i Un. STAT1 signal transducer and activators of transcription 1. Ch. SUMO Small ubiquitin-like modifier. engchi. TEMED tetramethylethylenediamine Thr (T) threonine Trp (W) tryptophan Ubc9 ubiquitin-conjugating 9 WRPW Trp-Arg-Pro-Trp. IX. v.
(111) 政 治 大. $& 學. ‧ 國. 立. ‧. -0 n. er. io. sit. y. Nat. al. Ch. engchi. 1. i Un. v.
(112) ɸ ʜͭγ ɸ ̝ϊʁͯȖό
(113) ϊʁͯȖόşļ̠ ͎"щÏē ϊʁͯȖό(learning and memory) ƓʢƿƺşZʻƟȵ̼ˎ VĨɯr "Ėʃ¸̱ȘйơşŢ̬̍ ʰnƓˈǬˈĪʗ¸ϊʁɎİ̋ĜŝşīϘ ϊʁƓ;Εȇ̞ ďɻɎİ̇Ľ˹ĤşȘǦϯ˝Ǭ˱P̞ѪşɿϛÏʒ͜āѧÔƠşQϹÏȖόŻƓŶȁϊ ʁşÂŏ ɂɎÃĬşȘǦϵ½Pª¶ό(recall) şϓĭ ÈϓĭşƿƺѸŊ̚ªÇ̜Ĭ ¯ΔŞŗşǬNj ȓȿѭNJѫĘʑȯvη̚şQϹ ÏАã+ Çşɕńϫªˋ µ È ϊʁͯȖόĦŤşчŸƨ̚ɀJ ź7š ;Εȇ̞ďɻ|ɂɎÃĬȘǦIå¸:¯ ̥ȶ;ˬÔʃɮ ģ»ţ˱ϊʁȖόͯǻȩȟ (hippocampus) Æч(Morris et al.,. 政 治 大. 1996)ǥљϊʁȖόͯą@ǯ(amygdala) Æч(Bear et al., 2001c) ˉÏ ϊʁͯȖόƓ NJήвşϔˎ ϯ˝ūrƪĎ6ħPI4ŝЮϊş÷ʑˬǞ ȧ ̼{ņɍȗşǕϊͯ šZɮϊşƪĎ ūÀĤƫϊǛˬ ćɈɽƵFȼµͯȘǦβɌş^±˝[ ̯͊Ђ̭ʎ NJήвşϓĭ. 立. ‧ 國. 學. !
(114) ϊʁͯȖόş͢щ. ‧. Ȗ ό õ Â ş ̼ ˎ y í NJ ˲ ƙ Χ ђ (encoding)
(115) ϵ ½ (storage)
(116) м ĵ. y. Nat. (consolidation)
(117) ʷį(retrieval) ;Εȇ̞ďɻɎİĤÐ̇Ľďɻş°͢ȘǦƈ ¸3̥ɂ. sit. n. al. er. io. ȘǦЭʸÂǿǵşΧђ Ơ ϵ½şƺ ɂȖόȯ°͢:¯şЭʸϵ½ÑȖόďɻ ̚ª Ç̻Ĭƨ¯şŞŗǬ ϵ½¸3̥FşȖόƿ̋ʇʷį njƷʎĝϿ̝;Ƥ٠NJvȸ е ʗ̍ġȖόʇϩö. Ch. engchi. i Un. v. Ä ʽ š Z ɮ ϊ Ǜ ɂ Ȗ ό ş ϔ ˎ ğ ώ ϵ ½ ¸ 3 ̥ Ǭ ˱ ş ŭ ˌ ȶ I Ơ è Ȗ ό (immediate memory)
(118) ˌʽȖό(short-term memory)
(119) ŭʽȖό(long-term memory) èȖόƠ̇ĽďɻİĬ ƥīϘƈņɇĬş®ˣ ȓƎўΒƬє ƷČÆǿåşȖØĚϵ ½¸3̥ŵƆ÷̍öȖ ˌʽȖό|oʔ̼ɽƵFǓÆş ĝʅδƟL¸ˌǬ˱Fāѧɽ Ƶ˱ş >Þ µÈˌʽȖό|oƎўΒIєÑΒ5Ǭ ŭʽȖόŻ|oƎўΒRĬΒNJ ` Ƥ Ñ ʀ Ĩ Ɉ ˌ ʽ Ȗ ό Ĭ ŭ ʽ Ȗ ό Ɠ ƺ ̞ ̼ Ȗ ό ş м ĵ ϔ ˎ (memory consolidation) ȷͯ -ʯƓņϡşȖόõÂ(memory formation) ̞ȇ̞Dƨчȼµş Ūɯ
(120) ʅδ³ÂoPȇ̞Ʈё˓͕şƿɾ(remodeling) ġȖόȓȿɈ NJˌΓ:мļş Ş͏ЭʸÂ̶ƠƎ,jмĵşȖό(Goelet et al., 1986 Squire and Alvarez, 1995) Ȗόȧ. ğώϵ½¸3̥Ǭ˱şŭˌȶI Ȗό|ªğώõÁş:¯IƠʛƾŃȖό. (declarative memory) (ͣƠ ѩ(explicit) oPŲʛƾŃȖό(nondeclarative memory) ( ͣƠFѩ(implicit) Ħщ ʛƾŃȖόјŊÆ̆тş¶ό̼{ņˋşĜ£ ģ»+Тƌ 2.
(121) ˫{ ǘ̬ ńŤƓƕRßƭ AΆÕō
(122) ´ AΆ ʎʗƓ|oēͷʵƾşȖό ˉÏ ÈȖόΘÁ̶ƠˌΓ ͎3ʖ¥ş"ÏēƆшȖɇ{ÀşƗ R{ ǘ̬
(123) ȯ A Ά Зˉ ŲʛƾŃȖόbȚʛƾŃȖόƓ̶шõÂş јŊŲ̆тşȖό Ȗόõ ƈ |oЭʸÂŭʽȖό ÈȖόΘÁyí(1) úʆŃȖόͯʁ͑ŃńúȓŃÆчşȖ ό ģ»ϊ̍. ƊΆ˰ė Ě:̍µƠƆ,ČÆ˰ėÏöȖ(2) ŷˋ(priming)̞̼ʷ. şȖό ģ»˖= NJ]¼ş¼DžèȓòϑƍvϑNJʩ¼(Warrington and Weiskrantz, 1968)(3) }ĩĭư(classical conditioning)ȵŝϊʁĬÆƗ͢īϘˋƈ Ɏ˝ɂ̍ª ˋ NJĜ£ µÏÆşQϹ ģ»øljʗš̹ʉ¹ʄÆƚ ¸ʘ Ɵȵ̻Ĭʄèȓȯ vͽбşQϹ(Milner et al., 1998) )
(124) ȷͯϊʁͯȖόş̥ȶ 1952 ÀPaul D. Maclean ʷĬфΨȇ̞ďɻ(limbic system) yíˈΒ¸3̥δ (cortex) P δ * ȶ Ȼ (sub-cortex) ş ˓ ͕ » * ˢ k (hypothalamus)
(125) ǻ ȩ ȟ. 政 治 大 ĝ̥ȶȷͯşxȓyíɊͭPţ˱ϊʁȖό(Maclean, 1952) ǻȩȟ¸фΨȇ̞ďɻþ͖ 立. (hippocampus)
(126) ą@ǯ(amygdala)
(127) фΨδ(limbic cortex)
(128) ;̾ȶ(septal area) ʎ. ‧ 國. 學. ʢlƺş˓͕ şxȓyí¹͢ȘǦş̄ϑPɰ̋şȖό. 1957 ÀScoville PMilner ˋŪşƪĎƏvHenry Molaison (H.M.) Ȃɉ¸ ̑ę À. ‧. µƠ̥ʖİĬ˼ǚÏϋƶѬѻ(epilepsy) ϯ˝ÀїΊŭÏѬѻȂȃʒ͜ʴL èġoпŝ. y. Nat. ŘЀ-ˈǩşɊŗ* ɂH.M. 3̥Ħфşѽ̧(Temporal lobe) FȲJȧ Ĩ;yí. H.M. şȂȃ ŽµÈϋƶ̋şȖόõÂİ̉. io. sit. ȩȟşʖ¥ ʎΔşŘЀ^Á σͥā˗. ǻ. er. ģ»H.M. ˈŖȖØå"͎qͺ̼ş̰Č¹,ʯ̍öȖ ʎ˵ˋɯDžƃу͌. al. ǻȩȟȷͯ. n. iv n C hengchi U ņÆşȖό ʔ̼ΰH.M.ʵох4+ş͊ǰ ̍ϯ˝ΦʁÇΒşΊw ѱ͍H.M.. Ȗόмĵş̼ˎ(Scoville and Milner, 1957) Milner ˑϊŤˋɯǻȩȟİ̉şȂɉĚŲ ʨ. ʵоşȓ&ʒ͜ˬć |đŲʛƾŃȖόĚČÆİĬΎѠ ĚjH.M. БȓȖɇ5ǬNjʼ̞ ˋşĜɊ -ŶÆͷēPúʆŃşϊʁȓ&ͯȖόşȓ& ʎΔş˓ŏѩǻȩȟ:Ɠŭ ʽȖόȹ ϵ½ş×̟ ǻȩȟƼʋʛƾŃȖόşõ Ě:ΎѠŲʛƾŃȖό µÈ Ȗ όĚŲ ͢õÁ :¯şȖόõÁ:¯̥ȶƼʋȽÔPʃɮ (Maclean, 1952) ą@ǯlƺͯɊͭP͇ʴǚłşϊʁȖόÆч(Amunts et al., 2005) јŊŲʛƾŃȖ όşǥљĭưϊʁ(fear conditioning learning) şȖόͯą@ǯÆч ˉÏą@ǯĚŲͯ ŲʛƾŃȖόÆч -yí. µɊͭşΎѠÏwɆ. ȖόşõÂşʛƾŃȖό(McGaugh et. al., 1996) ЩxȓŃǯ͟©ǧ(functional magnetic resonance imaging, fMRI) şƪĎÏ ˋɯúʆŃϊʁȖόͯȊŞѫ(striatum) Æч »XŬʿdȃ(Parkinson’s disease) ɉŤ ŊȊŞѫȞLÏϋƶĨúʆŃϊʁȖό̶ǟ(Sarazin et al., 2002). 3.
(129) ɸ!̝
(130) ţ˱ϊʁȖόͯǻȩȟɾФ
(131) ǻȩȟş͕ʓͯÿǝ̴Ǥ ǻȩȟ¸ŭ,oĤ Šʇļͯţ˱ϊʁȖόşõÂÆч ǻȩȟ¸ˋĐ̼ˎ;Ɠ̞ ѽ̧FȲȶȻ±ʞ̧МǞÏ ׸Ȳ̥ƁѽĒ(temporal horn of lateral ventricle) Ŀʖ Ơ~z̥͎ͣş˓͕ ǻȩȟŊŹŞJǂ*êC ¼ƀ µ ƀ+ͅHÌşĒ(ʇͣƠů Ͱ(Ammon) dĒ(Ammon’s horn cornu ammonisУͣCA) ǻȩȟ¸˓͕+ŠɎń˱ Ɏşͯ3̥δşʉ¹ȶȻPĨqфΨďɻ˓͕ʗÆʐɎ ǻȩȟή³ѫ(hippocampal complex) ˓ ͕ ǻ ȩ ȟ (hippocampus)
(132) ς Ş ȟ (dentate gyrus)
(133) ǻ ȩ ǫ ȟ (parahippocampal gyrus) ɾÂ ǻȩȟşδɾФ|IÂ)ΌI4Ό (molecular layer)
(134) ϮŞɽƵΌ(pyramidal. 政 治 大. layer)
(135) ¹õɽƵΌ(polymorphic cell layer) Ĩ;ϮŞɽƵΌşϮŞɽƵ(pyramidal cell) ƓǻȩȟʢlƺşɽƵ(Amaral and Witter, 1989) ğώϮŞɽƵõ͏PɎİÿǝѦͬş:. 立. ¯|ɽIƠCA1
(136) CA2
(137) CA3 ĨɽƵ35ƠCA3 > CA2 > CA1 (Bartesaghi and Ravasi,. ‧ 國. 學. 1999) CA1 ȇ̞ɽƵoϮŞɽƵƠl Ĉ ΌϮŞɽƵíÆ3 Ñ6 NJȇ̞ɽƵͮ˴¸Ĩ ; ϮŞɽƵÆĦ͢ϒƮ(dendrite) ͢ƓȼϒƮ(basal dendrite) ͢ƓʞͤϒƮ. ‧. (apical dendrite) ϮŞȇ̞ɽƵ<ņo(ʇͣƠ¹̐ȇ̞DɽƵ(multipolar neuron) Ɠµ ƠȼϒƮʏɃíÆĦNJ:¯^±ÏŀÛvɽƵѫ(cell body) şϒƮ (Bartesaghi and. y. Nat. sit. Ravasi, 1999) ʞͤϒƮşϒƮŭƃ̶ŭ б˰ϮŞȇ̞ɽƵƈ̍ʾ±ǻȩȟŀÛ µ. er. io. È È͢ϒƮlƺɎİĤÐCA3 şȇ̞ȘǦ (Bartesaghi and Ravasi, 1999) È ςŞ. al. iv n C molecular layer)
(138) ¹õɽƵΌ(polymorphic layer) ςŞȟlƺşȇ̞ɽƵƠИɺɽƵ h e n cell hi U c g (granule cell layer) Ĉ ΌИɺɽƵΌ4 Ñ6 NJɽƵͮ˴Ï ¸ςŞȟşȇ̞ɽƵ< n. ȟ - Ɠ I  ) Ό ɽ Ƶ Ό И ɺ ɽ Ƶ Ό (granule cell layer)
(139) Ç ɽ Ƶ I 4 Ό (acellular. ϒƮјŊʩ ^± ƒ(ʇͣ<Ơʩ̐ɽƵȇ̞D(monopolar neuron) (Johnston and Amaral, 1998) ǻȩȟȇ̞ȘǦ˹;ʢlƺIÂ)ɠ̴ǤʌƭѦ̴ͬǤ(Perforant fiber pathway)
(140) ƹŞѦ̴ͬǤ(Mossy fiber pathway)
(141) Schaffer collateral ʌƭѦ̴ͬǤƓǻȩȟϥ#ȇ ̞;ʇƪĎʢƠɤ̏ş ɠ̴Ǥ Dž§F·δΌ(entorhinal cortex) ЩÔÁÿǝ (longitudinal projection) şƭʔѦ̴ͬǤɂɎÃĬşȘǦ˹ĬςŞȟşИɺɽƵ ςŞȟş ИɺɽƵɂɎÃĬşȘǦ̄ϑ<ƈÏõÂMossy fiber ªɂȘǦ˹;ĬCA3 ȶȻşϮŞȇ ̞ɽƵ ƒʎɠ̴Ǥͣ<ƹŞѦ̴ͬǤ Schaffer collateral ŻƓɂCA3 ȶȻşϮŞȇ̞ɽ ƵɎÃCA1 ȶȻĬşȘǦ˹;ÑCA1 ȶȻşϮŞȇ̞ɽƵ Ï̚ȘǦ˹;ÑCA1 ȶȻ̍ ª̞ːЄ(fornix) ɂȘǦϥvÑǻȩȟ ʢƈ ņÆşȘǦ̞̼ǻȩȟ̄ϑʃɮ̼ªЩ 4.
(142) ϥvȇ̞DɂȘǦϥv˹;Ñ3̥δΌ
(143) F·δΌ
(144) *δΌ(subcortex) ϥvȇ̞ DoĦɠ̴ǤƠlIåƓ(1) ̞*̢(subiculum) ˹;ÑF·δΌªĬ3̥δΌ (2) ŠɎń˱ɎɂȘǦ˹;Ñ*δΌ(Amaral and Witter, 1989 Burwell et al., 1995) !
(145) ǻȩȟƨчşȵŝÔƠΘÁ ȧ ʔ̼ЈôȂģşƪĎ ƪĎ"Ǘ-ЩȵŝÔƠΘÁoѱ͍ǻȩȟ͎ŊȖόşΎ Ѡ ɃđşȵŝÔƠΘÁÆ(1) Ɋ͋Áĭưϊʁ(contexual fear-conditioning learning)(2) ʩ±ĀĭŃ̵Аϊʁ̯Ѫ(one-way inhibitory avoidance learning)(3) ʂdeȝƜ̯Ѫ (Morris water maze learning) 1998 ÀAtkins ɂ NJ;Ńĭưµ4(Ɋ͋)ͯ NJ͇ʴŃ ĭưµ4(̿̐īϘ) ˓³ˬÔĭưÔƠˇ̯ ëš͌Ѫȵŝ̚;Ńĭưµ4vɯǬ ª̼ :,ɂÆ NJ͇ʴŃĭưµ4(̿̐īϘ) şvɯ µÈ ̚;Ńĭưµ4ªƃvɯǬ Ʒ ͌ѪȵŝBȖɇ§źş̞Ѫ̍vɯǥљşQϹ ÈƠɊ͋Áĭưϊʁ(Atkins et al., 1998). 政 治 大. ʩ±ĀĭŃ̵Аϊʁ̯ѪŻЩÍ̈́şRŃƓʧºƇ¸˸̌ʃ ƒÈ̯ѪƓɂÍ̈́ʼn̟¸ NJ¯ǬħÆŌųȶP˸̌ȶşÔƠΤ; Í̈́ ˰ĺ̍ϨϏˬ#˸̌ȶ ÈǬ˖=̿̐ī Ϙ ̚*ǪɂÍ̈́ʼn#ÔƠΤǬ ƷƓ͌ѪȵŝBȖɇ§źʇ̿̐Ĭş̞Ѫ ŻϨϏȭȁ. 立. ‧ 國. 學. ŊŌųȶşǬ˱̶̍ŭ(Kesner and Hardy, 1983) Ɋ͋Áĭưϊʁͯʩ±ĀĭŃ̵Аϊʁ ̯ѪʎĦ͢ȵŝÔƠΘÁ̯ѪƓĦĜ£ƨ>чʐşȖό ¸Υγ]ƪĎ;ņġşȵŝÔ ƠΘÁ̯ѪlƺƓɍȗţ˱ϊʁͯȖό ƒɒʂdeȝƜ̯Ѫ. ‧. y. Nat. 1971 ÀO’Keefe ɂ ̿̐ʼn#3̈́ǻȩȟȶȻƯϭĨȇ̞̿× ˋɯǻȩȟFşȇ̞. sit. ɽƵ͎Ŋǿļţ˱×̟̍ɰȵÞ̿× ÏɂÈɰȵÞ̿×<ɽƵͣ<×̟ɽƵ(place. n. al. er. io. cells) (O’Keefe and Dostrovsky, 1971) Ï 1989 ÀMcNaughton ıFoster ƧʷĬ3̈́. v. ¸˸̌şϿ͋; ×̟ɽƵĚČÆǿåşƟȵ ̚˖= Ōųϙ¦̘ǝƈ×̟ɽƵ9̍Æǿ ǵşȳºŃ ǿåşƓ ƷƓ˖= Ōųϙ¦̘ǝƈϯèɂϙ¦чʙ ×̟ɽƵşȳºŃğ. Ch. engchi. i Un. Ц½¸ µÈ |oЂ̭Ĭ×̟ɽƵĚŲÆİǀŊ ƥşˢѐΩȌ ÏƓ̍ɂˢѐΩȌͯ ĨqşΩȌȯv NJţ˱Џ͊(spatial map) şʐ˓(Foster et al., 1989 McNaughton et al., 1989) 1984 ÀMorris ʷĬʂdeȝƜ̯ѪşȯŖƓɂÍ̈́ʼn̟Ŋ NJ:ʔŌş˅œÊ; ¸ eǂ*ʼn̟ NJàŊeǂΒHIş ÏϽψ+˖=ˢѐΩȌ(cue) ¤Í̈́ÐɍȌ oʮüЖЧŊeǂ*ş ̚ĨüĬĚ˅+ è|Ͼɇ¡Ǧ ЩÈeȝƜ̯Ѫ ƯϭÍ̈́ʼn#e;ŠĬüĬņŧ˥şǬ˱ ͣ<ȠбǬ˱(escape latency) ɐОÍ ̈́ʮüşȠбǬ˱˧ˌŪĨţ˱ϊʁȖό̶Ġ(Morris, 1984) Ŋ ʂdeȝƜ̯ ѪĨţ˱ϊʁȖόşȼСƓƄ¸ ¸Ͽ͋şȷÎŝ(reference) (ͣ<eȝƜȷÎǢΖ Ȗό̯Ѫ(water maze reference memory task) 1990 ÀEichenbaum ɡЂ̭ƓéȆзǻ ȩȟ̍ΎѠĬţ˱ϊʁȖό §ɂ3̈́şǻȩȟȆзƈªˬÔʂdeȝƜ̯Ѫ ƷƓĈÇ ̯ѪşˬÔʗɂ3̈́ʼn̟Ĭƨ¯ş×̟șΦĨoĵļ^ÁʮüЖЧŊeǂ*ş ѱ͍ 5.
(146) Ĭ3̈́ȓȿoĵļş^ÁΡşʮüĬˉÏ ̚ĈÇ̯ѪşˬÔʗϯϓşɂ3̈́ ʼn̟Ĭ:¯ş×̟ńƓˆUϽψ+şΩȌ ѱ͍Ĭ3̈́ʯˈŖЩϽψ+şΩȌͯΖņ ¸·şƨ͎ţ˱×̟şчŸȯvΡşʐ˓ ÏˈŖõÂţ˱ͯ͊(Eichenbaum et al. , 1990) È|oЂ̭Ĭǻȩȟǿǵş˓͕Pϫͨήвşȇ̞ȟ̴ µÏȓɂ:¯şȘǦʔ ̼ȇ̞ɽƵ˹#ǻȩȟ ̞̼ʃɮƈɂȘǦª˹Ĭ:¯ş̥ȶ µÈǻȩȟ¸ϊʁͯȖόş õÂ̼ˎþ͖ƿƺşĒÒ. ɸ)̝
(147) Hes-1 ʅδCɼ
(148) Hes-1 ȼµ˓͕ͯЭϭƟŃ Hairy and Enhancer of split 1 (Hes-1) ȼµјŊǖěщŏр¯̓şHairy and al., 1992. 治 政 Sasai et al., 1992) Hes-1 јŊȼƀЋəϿЋə(basic 大 helix-loop-helix, bHLH) şЭϭ 立 Āĭ4 (Akazawa et al., 1992 Kageyama et al.,1999 Kageyama et al.,2005. Enhancer of split (Hes) Ǜɘ lƺşxȓƓβ̝ȇ̞şˋĐ(Akazawa et. ‧ 國. 學. Kageyama et al., 2008) Hes-1 јŊHes Ǜɘ 3ÂǗ< Hes-1 ıHes-4 ˓͕+̶ ƨÜŊhairy ÏĨqÂǗş˓͕ŻͯE (spl) ̶ƨÜ -Æ ĝHes ƨчşbHLH ȼµ. ‧. ģ»Hesr/Hey/HRT/Herp/CHF/Gridlock (Iso et al., 2001) ıHes-like (Miyoshi et al., 2004) ̍õÂ:¯şĞƀǛɘ ¸Hes ǛɘÂǗ; Hes-1
(149) Hes-3
(150) Hes-5¸Í̈́şȇ̞. y. Nat. sit. ˋĐ̼ˎ;̍ȫƃŪɯ¸Ĩȇ̞̃ɽƵ (Bae et al., 2000) ̞ż^ρЛŖ(Southern blot). er. io. Pŝ͢˱ 0b͎(interspecies backcross) Iőу͌Í̈́şHes ȼµƓ NJʩ ή͵. al. iv n C ĚjЩS ǯ(S nuclease) PY4ŀÛ(primer ͌ѪüvHes-1 ȼµũĺЭ h e n g c hextension) i U ϭ Þ × ̟ (transcription initiation site) Ɠ × ¸ TATA ɵ 4 * ˅ ş ɸ 31 NJ ǯ n. (single-copy) şȼµ ×Ŋɸ16 NJƖÒѫ+şɸ26 NJ×̟ (Takebayashi et al., 1994). (nucleotide) (Takebayashi et al., 1994) Ĉ NJHes ȼµʗħÆ)NJconserved domains (1) bHLH domain [yíѯŃȶȻ (basic region) PЋəϿЋə(helix-loop-helix, bHLH)] (2) Orange domain ( the helix 3– helix 4 domain) (3) WRPW ( Trp-Arg-Pro-Trp ) domain ʎ͎ŊЭϭƟŃşβ̝ƓƆƿƺ ş(űϭ) (Kageyama et al., 2008) bHLH domain ×ŊѴȼͤȶȻ ͎Ŋ!ͮѫşõÂdimer formation) oPͯ deoxyribonucleic acid (DNA) ˓³ƓƆƿƺş ȼƀЋəϿЋəµ4ʔ̼bHLH domain ;şȼȶȻͯΖȼµ˓³Ěj¸ bHLH domain;ş HLH domain õ¯ƀͮѫ (homodimer) ńŤɳƀͮѫ(heterodimer) (Sasai et al., 1992) ǿåşƓ ¸bHLH ȼµ; 6.
(151) şHes ȼµĨbHLH domain şȼȶȻş;˱ʗħÆ NJȫƃŶȁşproline (P) Ějу ͌proline |ȓͯǿåşΖDNA ó¬ȫƃ˓³ȓ&Æч (Sasai et al., 1992) Æ]эƏv Hes-1˓³ĬN ɵ4(N boxCACNAG) ıĩƀşC site (CACGCG) şϠıŃb˓³ĬE ɵ4 (E boxCANNTG) ȫ ĚƏv̚ȆзHes-1 şN ɵ4̍ġĨĀĭÞİĬъƿş̉ ǚ(Sasai et al., 1992) µÈ Hes-1 |ʔ̼˓³ĬN ɵ4ÏĀĭјŊȼƀЋəϿЋə şȇ̞Ńȼµ»Mash1 Ūɯ ˉÏ ͎ĨşƟLƀȼƀЋəϿЋəȼµÏē Ż͎ Ŋ˓³ĬE ɵ4şϠıŃ̶ȫ(Chen et al., 1997 Takebayashi et al., 1994 Sasai et al., 1992) Orange domain ׸bHLH domain ş*˅ 7у͌Orange domain íÆĦNJг̐Ń Ћə(amphipathic helices) Ě7šOrange domain ͯbHLH ȼµņŪɯ<ʅδͯʅ δ˱ɰş >ÞÆч(Dawson et al., 1995; Taelman et al., 2004) ģ»Hes ƨчş. 政 治 大 ƨчşȼƀЋəϿЋəȼµE. ȼƀЋəϿЋəȼµHairy |ЩOrange domain ıȼƀЋəϿЋəȼµScute (spl) ɰ >Þ 立 al., 1995) ^ǂ Orange domain -ȓβ̝ЭϭĀĭ(Castella et al.,. ɰ >Þ ˉÏŽ:ȓͯHes. 學. 2000). ‧. ‧ 國. (Dawson et. WRPW domain ş×̟νūŊ C ͤ (carboxyl terminus) ńƓ׸C ͤ ʎNJdomain domain) WRPW. domain ̍ͯ©¯Āĭ4(co-. y. Nat. lƺÞƠĀĭ˓͕Ȼ(repression. sit. repressor) TLE (transducin-like enhancer of split) / Grg (a homologue of. Drosophila. er. io. Groucho ) õ >Þ ĚЩ˴ɾʅ{ Ѽ(histone deacetylase, Rpd3) ÏǍ͂Ɩ. al. n. iv n C al., 1994 ; Fisher ̍Щ˴ɾФʅ h e netg al., i U c h1996)Ĩ; Grg. Ò δ (chromatin) ş ˓ ͕ µ Ï ġ Ɩ Ò δ : Ɵ L Ě j Ɵ L Э ϭ Ā ĭ Þ Ɵ Ń (active repression)( Paroush et. { Ѽ(histone deacetylaseRpd3) ÏǍ͂ƖÒδ (chromatin)ş˓͕ µÏƟLЭϭ ĀĭƟŃ(represses transcription) ͣ<active repression (Chen et al., 1997). Ŋ Hes-1 ȼµ|ƟLЭϭĀĭÞµÈʇПщƠĀĭ4ƀşȼƀЋəϿЋəşȼ µ ЩЭϭÞşIőƏvHes-1 ȼµþ͖Ƽ±β̝Ť(negative regulatory) Ɠʔ̼Ħ͢ :¯şϓĭ(1) ŠɎ˓³ĬΖȼµşN ɵ4ÏɰĀĭÞP(2) АãĨqşƟL4˓ ³ĬE ɵ4 (Sasai et al., 1992) !
(152) Hes-1 ȼµŪɯşȘ̩βɌ Hes-1 ȼµŪɯşlƺİĬNotch. signaling şβɌ NotchƓ ̳͢Ϋʅ. (transmembrane protein) |oЩͯlƺşligand ˓³»Delta
(153) Jagged ņƟL Ɏ ˝Щγ I̭(γ-secretase) Jʣ ġNotch ɽƵFȶȻ (intracellular domain, ICD)Ɉɽ 7.
(154) ƵΫЭɷĬǯF ĚͯDNA ˓³ʅδRBP-J õ NJЭϭƟL4(transcriptional activator) ή³ŝÏͻϋHes-1 ıHes-5 şŪɯ ˉÏ ̚ICD ͯRBP-J ČÆõÂή³ŝ şǬNj RBP-J ħÆЭϭĀĭ4şĒÒ̍˓³Ĭ Hes-1 ıHes-5 ɖȵ4+ÏĀĭljşŪ ɯ (Artavanis-Tsakonas et al., 1999 Honjo, 1996) Æ]эƏvNotch |Āĭȇ̞ILPЩĀĭȇ̞̋(neurogenesis) ͬƎȇ̞̃ɽ Ƶ(neuron stem cells, NSCs) (Gaiano et al., 2000) Ƌļ̚ȐU Hes-1 ıHes-5 Notch ŻˈŖĀĭȇ̞ILPĀĭȇ̞̋ ʎrŪ˝Hes-1 ıHes-5 ¸Notch signaling ;þ͖ƿƺşĒÒ(Ohtsuka et al., 1999)¸;Εȇ̞ďɻˋĐ̼ˎ Hes-1 ıHes-5 Þ ƠNotch ş*˅ lƺŪɯ¸ventricular zone (VZ) þ͖ȇ̞ILşƼ±β̝ŤPŷˬȇ̞ ̃ɽƵǿŃ(Ohtsuka et al., 1999) signaling βɌ ȧÈ< ¸Äʽ 政 治 大 Hes-1 şŪɯ̍ˋ¸Notch ıDelta БŪɯ<ź ʎrŪş̠̆ƓNotch signaling Ě 立 ŲlƺΎѠHes ȼµŪɯşβ̝Ť ¸ʉ¹şƪĎ; Əv Shh (sonic hedgehog)
(155) Wnt. 學. ‧ 國. ˉÏ ĚČÆуώƏvHes-3 Ūɯ̍İĬNotch. signaling ıBMP (bone morphogenic protein) ¯Δ-ȓͻϋHes ȼµŪɯ µÈ Hes signaling βɌ-İĬĨqȘ̴̩ǤβɌ(Issac et al., 1998. Nat. )
(156) Hes-1 ȼµ¸ȇ̞ˋĐďɻşŪɯ. io. ȇ̞ˋĐ̼ˎ; ȇ̞+ɽƵ(neuroepithelial. al. sit. y. ‧. Nakashima et al., 2001 Solecki et al., 2001). cell) ˰ĺ̍õÂȇ̞Ő(neural. er. ȼµŪɯȧ İĬNotch. n. iv n C 2004) +ƾʷĬHes-1 h e n g c h i Uȼµ|Āĭȇ̞IL Æ]эƏv. plate) ıȇ̞ͥ (neural tube) Њψ ʎĝɽƵʏɃ͖̍LÂϣǝŞȇ̞άɽƵ(radial glial cells) (Hatakeyama et al.,. Hes-1 ȼµƮѧş5̈́Ĩȇ̞IL̍¸źѢɽƵΊ˃ź§ÂΜ ÏʓÂȇ̞ͥõÂİ̉ ġ. ƳƴˋĐ̼ˎ;ńŤƓvƈČ¹,ϯèÉ/ (Ishibashi et al., 1995) ¸ȇ̞ˋĐ̼ˎş Äʽ Hes-1 ȼµ|oͬнȇ̞+ɽƵ(neuroepithelial cell) ıϣǝŞȇ̞άɽƵ(radial glial cells) şΒˋĐƈʽHes-1 ȼµ|oŷˬƔŞȇ̞άɽƵ (astrocytes) õ Hes-1 ıHes-5 ȷͯϣǝŞȇ̞άɽƵÄʽˋк ĦŤêɯ>̪şчŸ ЉģĤ ͺHes-5 ȫƃŪɯ¸;̥(midbrain)
(157) ƈ̥(hindbrain) Ï:Ūɯ¸isthmus ÏHes-1 ŻŪɯ¸isthmus (Hatakeyama et al., 2004) ƷƓ¸̥̈́ƳƴˋĐşɸ13.5 R Hes-1 ń ƓHes-5 ƷˈŖŪɯŻ̍Āĭȇ̞D(neuron) şILĚjĀĭϣǝŞȇ̞άɽƵşΒ (Ohtsuka et al., 2001) ¸Hes Ʈѧ5̈́;proneural. bHLH ȼµ »Mash1 ıNgn2. (Neurog2) ̍ȫƃŪɯ ʎʇˢƠ ͢:ÂΜşȇ̞̋ (premature neurogenesis) (Hatakeyama et al., 2004)Hes-1 ȼµǒȧ5̈́(knock-out mice) ¸ƳƴˋĐ̼ˎ;Æˈ. 8.
(158) ̥ ş ɯ ˣ ɰ j proneural ȼ µ » Mash1 Ū ɯ ˭ Ί w Ě j ȇ ̞ ̋ ̍ w ʑ ˬ Ô (Ishibashi et al., 1995 Cau et al., 2000) ¸ȇ̞ˋĐ̼ˎşƈʽ ̚ȇ̞άɽƵ̋(gliogenesis) Hes-1 ЩƔŞȇ̞άɽƵ ƎўŪɯ¸ˋĐş3̥; ȮġȐmHes-1 PHes-5 ʎĦNJȼµ ̍ŷˬ3̥FşƔŞȇ ̞άɽƵɰPˢͪΫ(retina) şMuller ȇ̞άɽƵɰ(Nakashima et al., 2001) ˉ Ï ̚ȐmHes-1 ńHes-5 ʎĦNJȼµĨ;< Ż̍ġMuller ȇ̞άɽƵɰˆU (Hojo et al., 2000 Nakashima et al., 2001) È|š Hes-1 ȼµ¸ˋĐ̼ˎş:¯Ǭ˱ þ͖:¯şĒÒ ¸Äʽ|oͬнȇ̞̃ɽƵşΒ˭ϸƈʽŻŷˬȇ̞άɽƵ̋ ¸ 2007 À Lee ˑ " Ə v ɸ ƀ Ó ɤ P Ϝ δ ȋ ͻ ϋ Ϙ Serum- and glucocorticoid-inducible kinase 1, SGK1)|ʔ̼Āĭͯ Hes-1 ʅδ¯ NJǛɘ;ş. 政 治 大. Hes-5ˬÏΊɆǥљϊʁͯȖό(Lee et al., 2007) ̼{ÆƪĎƏv Āĭγ I̭ (γ-secretase) ġɇ NotchİѫˈŖòÂJʣÞ ÏĀĭNotchȘǦ˹; ˬÏŷˬǥ. 立. љϊʁͯȖόoPţ˱ϊʁͯȖό (Dash et al., 2005) ź7šNotch*˅ȼµ Hes-5Ě. ‧ 國. 學. :ȷͯţ˱ϊʁͯȖό(Lee et al., 2007) µÈ NJ Notch*˅ȼµ Hes-1 |ȓȷͯţ ˱ϊʁͯȖόşõ µÈ Υγ]ƪĎ̈ˬ ćɍȗHes-1 ͯţ˱ϊʁͯȖόşƨч. Nat. y. ΎѠȇ̞ďɻ -ΎѠĨqɾФşɽƵILPĨɽƵõ͏ »Óɭɽ. sit. Hes-1 ȼµȧ. ‧. Ń. er. io. Ƶ
(159) FIŔ(endocrine) IŔ(exocrine) ɽƵ
(160) ѫ̝(somite) ˑ(Kageyama et al., 2005) ¸T ɽƵˋĐ̼ˎ; Ȑm Hes-1 ȼµ̍ġT ɽƵˋĐȭȁ¸Äʽ ÆεşƓƷ. n. al. Ch. i Un. Ȑm Hes-1 ȼµĚ:ΎѠB ɽƵˋĐ (Tomita et al., 1999). engchi. v. ɸ̝
(161) щ5ŚȋLǍ͂
(162) щ5Śȋ˓͕ͯIщ щ5Śȋ(SUMO) ¨²Ɠsmall ubiquitin-like modifier щ5Śȋʎ͢ʅδIƆ ½¸Ŋ°͢ȅǯŝ; ʢÄƓ¸Ϳ˞ S. cerevisiae ˋɯş Ĵ²Ơ SMT3 (Suppressor of MIF Two 3 protein) (Meluh and Koshland, 1995) щ5ŚȋɈͿ˞Ĭǖě щɽƵƧħÆȫƃşŶȁŃ(Kerscher et al.,2006 Ŋщ5ŚȋĨȼµó¬+3ưÆ18 % ͯŚȋ (ubiquitin) ƨÜj˓͕+ͯŚȋÆƨ¯ş)ͬ˓͕ (Hershko et al., 1998. Ohsumi, 1999) yƐ NJβ ͔Ѥ(β-sheet) ѝХ NJα Ћə(α-helix) şɭŞ͔Ѥ (Bayer et al., 1998 )(µĨʅδÞ^ÁщÜŊŚȋ ƒПщŊŚȋƨчʅδ(ubiquitin9.
(163) related protein) ş щ ȧÈ< щ5ŚȋͯŚȋĦŤ¸C ͤʗÆĦNJglycine ˂ȼ (di-glycine) ͯΖʅδşlysine ×̟õ©ΉщɳѳД(isopeptide bond) şʐ˓Ïȷ ͯQϹ(Gill, 2004) :¯şƓ щ5ŚȋşN ͤ(N-terminal)ưÆ10-25 NJŲ˓͕ŀÛ şѴȼ ʎNJ¸ĨqşŚȋƨчʅδ;ĚČÆˋɯĬ(Johnson, 2004) щ5Śȋ3ư100 NJѴȼņɾÂş I4˭3ưƓ10 kDa ş5I4ʅ (Bayer et al. , 1998) ̞ȼµó¬Iő Ϳ˞
(164) ŏрŪɯʩ NJщ5Śȋȼµ ǖěщ|Ū ɯNJщ5Śȋȼµ ÏˀŝĂ|Ūɯȫ̺$NJщ5Śȋȼµ(Kurepa et al., 2003) ź Ɉǖěщȼµó¬+ΧvNJ:¯Ğƀşщ5Śȋ IåƓSUMO1 (Ubl1, Smt3c, Sentrin, PIC1, GMP1)SUMO2 (Smt3a, Sentrin2)SUMO3 (Smt3b, Sentrin3)SUMO4 (Melchior et al., 2000)SUMO2 ıSUMO3 ƨÜŃ̐ȫ ¸ó¬+ȫ̺97 Ɠƨ¯ş ņoɃʇПщƠSUMO2/3 SUMO2/3 ͯSUMO1 şƨÜŃưÆ50 ÏͯSUMO4. 政 治 大 ÆşɾФ; ˉÏ ͎SUMO4 Ïē ŽlƺŪɯ¸˚ѥ
(165) ˙ѥPɥX˓ (Melchior et al., 立 2000) ƪĎSUMO ¸ɽƵFş×̟ ƏvSUMO2/3 lƺ׸ǯδ(nucleoplasm) Ï ŻÆ87 ƨÜŃ (Melchior et al. , 2000) ¸ˋĐş̼ˎ; SUMO1 - 3 ̍ŚŪɯ¸ņ. ‧ 國. 學. SUMO1 Ż×¸ǯΫ(nuclear envelope)
(166) ǯ@(nucleolus) (Ayaydin et al., 2004). ‧. щ5ŚȋņǍ͂şΖʅδ3¹yí NJ ƶŃó¬ ΨKXE Ĩ;Ψ rŪ NJħ. y. Nat. ÆɴeŃşѴȼ K Ɠ̍ʇщ5ŚȋǍ͂şlysine (K) X ŻƓ¤Ù NJѴȼ E Ɠ. sit. glutamic acid (Rodriguez et al., 2001) ΨKXE motif ͎ŊŠɎ˓³Ĭщ5ŚȋE2 ˓³. er. io. (E2 conjugating enzyme) Ubc9 (ubiquitin-conjugating 9) +ƓƆƿƺş Ějщ5ŚȋņǍ. al. n. iv n C ×Ě:¸ʎNJ ƶŃó¬; ź 7ΡļÆĦNJ:¯şmotif ģ»phosphorylationhengchi U ͂şΖʅδlƺş˓³×ƓЩlysine ʎNJ×̟ ˉÏ ÆĝΖʅδşǍ͂˓³. dependent SUMO motif (PDSM) ı negatively charged amino acid dependent SUMO motif (NDSM) (Wilkinson and. Henley et al., 2010) PDSM Ɠ NJSUMO motif. ψKXEXXSP ɾÂS rŪ Serine ʇЃL Ï P rŪƨθ proline ЃLş×̟ ūʽş ƪĎ˓ŏƏv¸in virto ńƓin vivo ; PDSM ЃL|oΊwщ5ŚȋE2 ˓³ş˓ ³PǍ͂ lysine NJ5Śȋ motifͣ<Ơ NDSM Ɠ NJ|ŀÛŃş motif ψKXEXXD / E ņɾ -ʇˋɯ¸Ĩqşİδ;þ͖ƨÜşĒÒjȓΡş̀ˇvщ5 ŚȋΖʅ ˉÏ чŊʎĝŲ ƶŃşǍ͂˓³×Ɠ»ÙʇǍ͂şźşƪĎľ:ɤ ̏ !
(167) щ5Śȋ|ȜŃǍ͂ʕǤ щ5ŚȋLǍ͂ƓЩ ď¬ş ġщ5ŚȋȓȿǍ͂Ζʅδ+şlysine(Hay et al., 2005) щ5ŚȋLǍ͂ʐ˓ʕǤyíÆ)NJlƺşćѮƟL (activation) ˓³ 10.
(168) (conjugation) ʐɎ (ligation) ljIåƓЩE1
(169) E2 ıE3 ʎ)͢:¯ş˻Lş(Liu et al., 2008) ˰ĺŪɯşщ5ŚȋƓ NJ:ÂΜşźѢʅδ(immature proform) ͤɄÆ ƙ ư2~11 NJѴȼ:ˑşˌц ʔ̼SENPs (sentrin-specific proteases) ¸ĨC ͤˬ ÔJȧ Ï<ƈ¸C ͤџvĦNJglycine ˂ȼ(di-glycine) 9ȓÂƠÂΜşщ5Śȋƀ Á µÈ9ȓˬ#щ5ŚȋLǍ͂ʕǤ(Bailey et al., 2004 Li and Hochstrssser, 2003) Dž§ ÂΜşщ5Śȋ̍ЩATP ʷĢȓ˭ġC ͤѶȼɰ̤L ˉƈͯE1 ƟL (E1 activating enzyme) şcysteine QϹƈ ѓʼnvAMP ġщ5ŚȋͯE1 õÂɶѸД (thioester bond) ©Ή˓³¸ Ț E1 ƟL¸ǖěщlƺşõÁƓSAE1 / SAE2 ņõ Âşɳƀͮѫ(heterodimer) (Okuma et al., 1999) Ɏ˝ щ5Śȋ̍ɈE1 ЭɷĬħÆ cysteine ƟL׺E2 ˓³+ ¯Δşͯщ5ŚȋõÂɶѸД ʢƈ NJćѮƓʔ̼щ5. 政 治 大 ȋşΖʅδ+ µƠ¸Ζʅδşlysine \цıC ͤş͠ȼɰ©ΉщɳѳД 立 (isopeptide bond) ʐ ˓ ş Ǎ ͂ Þ µ Ï ġ ɇ İ δ ş ͕ ƀ (conformation) ń Ɠ ˓ ͕. ŚȋE3 ʐɎ(E3 ligation enzyme) ˻L E2 ˓³(Ubc9) ̍ѭщ5ŚȋЭɷĬщ5Ś. ‧ 國. 學. (structure) āѧ ˬÏΎѠİδşxȓ(Gill, 2004) ¸Ɔ¹şɽƵѫ ͌ѪƪĎʗƏvщ5 ŚȋǍ͂Lƺʔ̼E1 ƟLıE2 ˓³ʯȓòÂǍ͂Þ ģ»¸ɽƵѫ . ‧. SUMO1 ͯRanGap1 ş˓³ƺʔ̼SAE ıUbc9 ȷͯQϹ èȓòÂǍ͂Þ (Hay,. y. Nat. 2005 Tatham et al., 2001) ÝƓ ɽƵF3¹Βşщ5ŚȋǍ͂LBƺʔ̼E3 ʐɎ. io. sit. şȷͯ E3 ʐɎЗˉ:̍ͯщ5Śȋ©Ή˓³¸ Ț ÝƓE3 ʐɎ|oͯE2 ˓. al. n. 2004). er. ³ıΖʅδ˓³¸ Ț ŷˬщ5ŚȋE2 ˓³ЭɷĬΖʅδ(Johnson,. Ch. engchi. i Un. v. щ5ŚȋLǍ͂Ɠ NJ|ȜŃȵ͏Ǎ̼͂ˎ(űϭ!) ¸ɽƵFÆ ͢ǿɳŃşʅ SENPs ȧ. ħÆC ͤḙxȓ(hydrolase) Бȓȿɂщ5ŚȋɈ7ˋǍ͂şΖʅ. +ɷȧ È̼ˎͣÞ{щ5ŚȋLǍ͂(de-sumoylation) (Schwienhorst et al., 2000) ¸Ϳ ˞;ͯSENPs ƨ͎Ϲş{щ5ŚȋLǍ͂şͿȋ IåƓUlp1 ıUlp2 (Gill, 2004) Ulp1 ׸ǯTή³ѫ(nuclearpore complex, NPC) + |Jȧщ5Śȋ immature ş ͤ ˌo P J ȧ щ 5 Ś ȋ ͯ Ζ ʅ ş Ɏ ³ Ulp2 × ¸ ǯ @ З ˉ : ȓ J ȧ щ 5 Ś ȋ immature ş ͤˌ ÝƓȓȿJȧ ĝǿǵşщ5ŚȋͯΖʅşɎ³ (Gill, 2004) ¸ǖěȵŝ; ŻÆ ͢{щ5ŚȋLǍ͂şͿȋ è+ƾņʷĬşSENPs q ljņÞşщ5ŚȋĞƀ-:¯ ĨɽƵFş×̟Pxȓ+-ɱÆǟɳ (Gill, 2004) Ŋ {щ5ŚȋLǍ͂şͿȋ:ȓȿġщ5ŚȋɈ:ÂΜşźѢʅδÂƠÂΜşщ5Ś ȋ -ħÆɂщ5ŚȋɈΖʅ+ɷȧşxȓ µÈ Æ {щ5ŚȋLǍ͂şͿȋ½ ¸ ɽƵFщ5ŚȋLǍ͂ş9ȓêɯƋļşŞ͏ 11.
(170) )
(171) щ5ŚȋLǍ͂şxȓ ЭђƈǍ͂Þ(post-translational modifications)yíȼL (methylation)
(172) ЃL (phosphorylation)
(173) ŚȋL(ubiquitination)
(174) щ5ŚȋLǍ͂(sumoylation) |ȜŃşЭђ ƈǍ͂ÞŚş̸¸ȵ͏β̝ʅδƟŃ ¸ɽƵȘǦ;˹;;-þÆƿƺşĒÒ щ 5ŚȋPĨǍ͂ÞΎѠΌǂƨ̚şŚ ̼{ƪĎˋɯщ5ŚȋǍ͂şΖʅlƺoǯ ʅ(nucleus protein) Ơl (Verger et al., 2003) ÝƓūÀĤşƪĎˋɯĬȷͯȘǦ˹;ş ʅ
(175) Эϭµ4
(176) Ȃƚʅ
(177) ͯDNA Ǎ̪ďɻƨч<ʅˑ-̍ˋщ5ŚȋLǍ͂ ʎ |oͺŌщ5ŚȋLǍ͂¸ɽƵǯ -ħÆ ļşƿƺŃ ̼{ÆƪĎƏvщ5ŚȋLǍ͂ |oβ̝Ζʅδ āѧΖʅδ<͕ƀ
(178) ƵFI×̟
(179) ЭϭƟŃ
(180) мļ ńƓΎѠ ͯĨʅδ<˱ş >Þ ȧÈ< -yíβ̝ɽƵFQϹ ģ»DNA ή͵
(181) ɽ Ƶ˫ʽβɌ
(182) DNA Ǎ̪Þ
(183) ɽƵǯ̸ϥÞ
(184) ƖÒѫIб
(185) ȼµѫòϑŃ
(186) β̝ЭϭÞ
(187) ȘǦ˹;
(188) ˋśQϹˑɯˣ(Bies et al., 2002 Gill et al., 2003 Kahyo et al., 2001. Nacerddine et al., 2005) . 立. ɽƵ˫ʽşβ̝. 政 治 大. ¸Ϳ˞; щ5ŚȋE1 ƟLıщ5ŚȋE2 ʐ˓şƮѧ̍ŰлɽƵ˫ʽ (cell. ‧ 國. 學. cycle) ˬÔj̍ʓÂG2 / M phase ȭ͛ġϑNJɽƵ˫ʽȭa(Seufert et al. ,1995)щ5Ś ȋǍ͂L͎ŊɽƵǂ͎ȼµƚŃϷ&(genotoxic stress) ş½Ɵ-Ɔƿƺ ¸Ϳ˞; Ā. ‧. ĭщ5ŚȋǍ͂L̍ΊwȼµƚŃşɗ̇Ѹ"щɽƵ; ŻˋɯĬ:ͥƓЩѩŃƼ±. y. Nat. (dominant-negative) Ubc9 ńƓµ̤Ȃƚ̇Ɩʔ̼ȂƚʅGam1 ̍ġщ5ŚȋE1 ƟL. sit. ǁ̭ÏĀĭщ5ŚȋLǍ͂ ʗ̍ΎѠɽƵ͎ŊȼµƚŃɗ̇ŃΊw ˬÏͻϋɽƵǐ/. n. al. er. io. (apoptosis) (Tempé et. al, 2008) !DNA Ǎ̪Þ. Ch. engchi. i Un. v. ƪĎƏvщ5Śȋ E3 ʐɎ Mms21 / Nse2 (Non-structural maintenance of chromosomes element 2 homolog) | o ġ Smc5 / 6 (structural-maintenance-ofchromosome, SMC) ή³ѫʔ̼щ5ŚȋǍ͂LǍʱDNA гťОˡ(Andrews et al., 2005 McDonald et al., 2003 Potts et al., 2005 Zhao et al., 2005) µÈ ƷƓʨE3 ʐɎxȓ̍ġ DNA ˼ǚɗ̇Ńʷȫ ¸DNA ή͵PǍ̪Þ;Æ NJƿƺşʅ proliferating-cell nuclear antigen (PCNA)¸ɽƵ˫ʽG2 ʽή͵ŊDNA ή͵ƈǍ̪̼ˎ ;þ͖ƿƺĒÒ PCNA +şlysine 164 ̍ˬÔʩ ŚȋL(mono-ubiquitination) ń¹Śȋ L (poly-ubiquitination)ˉϸS ʽщ5Śȋ̍ͯŚȋюŜPCNA +şlysine 164 ş˓³ × ŰОŚȋLˋ µÏĀĭDNA ˼ǚņͻϋşDNA Ǎ̪Þ(Hoege et al., 2002) È|Ђ̭ĬЗˉŚȋͯщ5ŚȋǍ͂Ζʅ¯ NJ×̟lysine ×̟ ÝĨΎѠΌǂŽ Ɠ:¯ş. 12.
(189) )ƵFI×̟ RanGAP Ɠɸ NJˋɯĬ|ʇщ5ŚȋǍ͂şΖʅ ͎ŊɽƵǯͯɽƵδ<˱ŝ δş̸ϥ(nucleocytoplasmic transport) ƨ̚ƿƺ RanGAP (Ran small GTPase activating protein) ¸ɽƵFş×̟Ɠ׸ɽƵδ ʇщ5ŚȋǍ͂ƈ RanGAP ̍ͮ˴ĬɽƵǯ фΨͯɽƵǯTή³ŝ(nuclearporecomplex) < şRanBP2 ˓³ oβ̝ɽƵFŝδ̸ ϥ(Mahajan et al., 1998). ʅδмļşβ̝ ƪĎˋɯ ɽƵFщ5Śȋ̍ͯŚȋюŜΖʅδşlysine ˓³× ˬÏŰaʅ δµщŚȋLÏˋǁ̭Þ ģ»¸ɃşɊŗ* ¸ɽƵδ;şЭϭµ4 NFkappaB ̍ışĀĭŝ Bα ˓³ÂŲƟL͏şή³ŝ ̚Æ ĤşīϘ»Tumor. 治 政 大 ̭ µÏġNF-kappaB ˬ#ɽƵǯFɖȵ*˅ȼµşЭϭ µÈ ̚ɽƵFщ5ŚȋLǍ 立 ͂ˋ |oͯŚȋюŜBα şlysine ˓³× Ïмļ Bα ͯNF-kappaB ş˓. necrosis factor (TNF) şÞ Bα ˋŚȋLÏʇ26S ʅѫ(26S proteasome) ǁ. ‧ 國. 學. ³ ˬÏĀĭNF-kappaB şxȓ(Desterro et al., 1998) ΥƪĎ¯Δ-ƏvŚȋLı щ5ŚȋǍ͂L̍юŜ¯ NJΖʅşlysine ˓³× ̚von Hippel-Lindau (VHL) ʇщ. ‧. 5ŚȋǍ͂|oβ̝ĨʅδşмļoPĨʅ¸ɽƵ;½¸ş×̟(Qiliang Cai et al.,. y. LǍ͂̍ΎѠĨмļ (Lin et al., 2003) щ5ŚȋLǍ͂ȧ. ŠɎΎѠĨΖʅşʅδ. sit. Nat. 2010) ¸Smad4 (mothers against decapentaplegic homolog 4) ;-Æѱ͍Ĭщ5Śȋ. er. io. мļ ¸2003 ÀGill şƪĎ;ʷĬSUMO1 |ƟLp53 şЭϭÞ Ějͯp53 ʅ˓. al. n. iv n C Protein 1 (Sp1) Ʈѧġ<:ȓˬÔщ5ŚȋLǍ͂̍bʘƀмļ Ăˬ ćƪĎѱ͍Ĭщ hengchi U ³ġp53 şʅδĂƠмļ (Gill et al., 2003) ΥƨчƪĎ ѩɂSpecificity. 5ŚȋǍ͂Sp1 |ΊwSp1 ˓³Ĭ26S ʅѫÏŷˬĨǁ̭ÞˬÔ(Wang et al.,. 2008) È|oЂ̭Ĭ щ5ŚȋǍ͂ΖʅÏΎѠĨмļşϓĭ -|ȓƓʔ̼Ĩq ϓĭªˬÏΎѠĨмļ ?ЭϭƟŃşβ̝ ¸ǖěȵŝ; щ5ŚȋǍ͂LşΖʅ3¹ƓЭϭβ̝µ4
(190) ©¯ƟLµ4ńƓ © ¯ Ā ĭ 4 ʉ ¹ ƪ Ď у ͌ щ 5 Ś ȋ Ǎ ͂ ş Э ϭ β ̝ µ 4 Ƀ đ ş Æ p53
(191) Elk-1
(192) STAT1
(193) C/EBPs
(194) SRF ˑ Э ϭ © ¯ β ̝ µ 4 Ƀ đ ş Ż Æ p300 P ˳ Ń Ϙ ȋ İ ѫ (androgen receptor) (Gross et al., 2001 Gross et al., 2004 Nishida et al., 2002) ȕÏ ē Эϭµ4ˋщ5ŚȋLǍ͂̍ĀĭljşΖȼµЭϭÞşˬÔ ˉÏ щ5Śȋ LǍ͂Ɠ»ÙĀĭȼµЭϭIJşϓĭ |ȓƓµƠщ5Śȋ˓³¸Ζʅ+ÏìY »HDAC
(195) Daxx ʎщşI4 ˬÏĀĭЭϭşˬÔ Æʉ¹şуώѩщ5ŚȋLǍ͂ |ȓ̍ʔ̼:¯ş^ÁĀĭ:¯şЭϭµ4ĨЭϭƟŃ Ĩ;ʢУʩş^ÁƓЩˆUЭϭ µ4şŪɯ˭ -|oЩ8НЭϭµ4ͯşΖDNA ó¬ş >ÞĀĭЭϭ 13.
(196) ƟŃ ģ»щ5ŚȋLǍ͂|oΎѠTFIID ˓³ĬşΖDNA ó¬ ʎ|ȓ̍ˆǣğϢ TFIID şũĺЭϭˬÔ щ5ŚȋLǍ͂ȧ. Ƽ±β̝ ̚Ν¡âʅ(heat shock factor,. HSF) HSF1
(197) HSF2 ˋщ5ŚȋLǍ͂|ˬÔ±β̝ ġ<ͯDNA ˓³ȓ&ΊɆ (Hong et al., 2001) È|đ щ5ŚȋLǍ͎͂ŊЭϭƟŃşβ̝Ɠ NJήвşϓĭ (Goodson et al., 2001 Johnson, 2004). ɸ?̝
(198) щ5ŚȋE3 ʐɎPIAS1 ʅδCɼ
(199) PIASǛɘCɼ PIAS (protein inhibitor of activated signal transducer and activators of transcription) ʅ δƓoSTAT1β (signal transducer and activators of transcription 1) Ơͻ΅ʅ ġ". 政 治 大 δ˓³ ̞͌ѪˋɯÆ 35ư650 立 NJѴȼşʅδ̍ıSTAT1. щB ɽƵǡ(B cell library) Ěʔ̼Ϳ˞гв ďɻIőƓéÆʅδ|ıSTAT1β ʅ ˓³ ɂ<Ĵ²Ơ. ‧ 國. 學. PIAS1 (Liu et al.,1998) ¸ § ź ş ƪ Ď Ə v æ © ¯ ã Ʀ Ċ ϕ Ŗ (coimmunoprecipitation)Iő¸in vivo Ɋŗ* STAT1 tyrosine 701 ×̟ˋЃL |ġ. ‧. PIAS1 ͯSTAT1 ɰ >Þ (Liu et al., 1998) Ăˬ ćƪĎƏv ̚İĬ8Нȋ (interferon) ΎѠ PIAS1 ̍ĀĭSTAT1 ņʫCşȼµŪɯ (Liu et al., 1998) ĚjЩɽ PIAS1 < ľÆ)NJʅ. io. er. (Liu et al., 1998) ̞ǖěщɽƵşȼµó¬Iő ˋɯȧ. sit. y. Nat. ƵϘȋşīϘ(cytokine) şīϘPIAS1 ıSTAT1 ˓³̍ǁàSTAT1 ͯ DNA ˓³şȓ& δĨѴȼó¬ɾÂͯ PIAS1şƨÜƃƆȫ IåƓ PIASx (ͣ<PIAS2)
(200) PIAS3ı. n. al. Ch. i Un. v. PIASy ((ͣ<PIAS4)ĚɂĨ ĥПщƠPIAS ʅδǛɘ (Chung et al., 1997 Copeland. engchi. et al., 1995) ȧ PIAS1 ĨqşPIAS ƧˋɯĬ¸mRNA ȴɎ̼ˎ;̍õÂĦNJɳõ ѫ(isoform) ǖěщPIAS ¯̓ʅ-½¸ŊŲȔˁȵŝ
(201) ˀŝıͿ˞ˋɯĬ "щş Zimp10 ıZimp7 oPͿ˞;şSIZ1 ıSIZ2 ¸ūÀĤ-ˋɯĬͯǖěщPIAS ʅħ Æȫƃ¯̓Ń (Johnson and Gupta, 2001) . !
(202) PIAS1 ˓͕Cɼͯщ5ŚȋLǍ͂şчŸ źşƨчƪĎ7šPIAS ̍Щ:¯ş˻L^ÁˬÏβɌ¹NJͯãƦƨчşЭϭµ4 şƟŃ ƒȓмļɽƵFãƦďɻşƋļ(Shuai et al., 2005) ΥƪĎlƺƺCɼşƓ PIAS1 ȧ. ÞƠSTAT1 şƼ±βɌŤ ¯ǬħÆщ5ŚȋE3 ʐɎşƟŃ |o͎. STAT1 ńŤƓщ5ŚȋşΖʅδˬÔǍ͂ (Liu et al., 2007 Song et al., 2006) ź 7šşщ5ŚȋE3 ʐɎyíÆPIAS Ǜɘ
(203) Pc2 ıRanBP2 (Jackson, 2001) Ĩ; ÆPc2 :ħÆϿŞ˓͕ ˉÏĨ;PIAS şƟLƺϿŞ˓͕ş½¸ PIAS ʅ̍ŷˬʉ 14.
(204) ¹şЭϭµ4ˋщ5ŚȋǍ͂L ÏāѧĨЭϭƟŃ ǖěщȵŝPIAS ʅ źˋ ɯĬ7šÆ ͢ lj¸Ѵȼó¬+ÑUÆ50 % Ɠƨ¯ş ̞Iő¸ljşN ͤı PƗĝȶȻ+ħÆȫƃŶȁŃ PIAS ş˓͕+ÆNJlƺşmotif N ͤSAP (scaffold attachment factor-A/B/acinus/PIAS) motifPINIT ( Pro - Ile - Asn - Ile - Thr) motif RING finger like zinc binding domain (RLD)C ͤȶȻʭíÆŃȶȻacid domain (AD) P serine ı threonine rich (S / T) ȶȻ (Rytinki et al., 2009)(űϭ)) ¸ʉ¹̍ͯƖÒδ˓³şʅ;ˋɯĬSAP motif ƪĎƏvPIAS1 ̍ıDNA ó¬+ scaffold attachment regions (SARs) Fʭí̤ѺѲ(A, adenine) ıȒ̤ѹѵ(T, thymidine) şȶȻ˓³ (Klapp et al., 1993) SAP motif íÆ ƙLXXLL (L Ɠlysine X Ɠ¤ÙşѴȼ Ŷȁó¬ ÆƪĎѩʎ͎Ŋǯδİѫ(nuclear receptor) ͯ©¯ƟL4(co-activator) ή³ŝşõÂƆƿƺ ȧÈ< ƷƓɂPIASy +şLXXLL ƈǂĦNJlysine ƮѧÂ. 政 治 大. alanine ̍ǁàPIASy Ǔ͎ŊSTAT1 ЭϭƟŃşĀĭ . 立. PINIT motif ½¸ņÆşPIAS ʅδ; PINIT motif ̍ͯRING finger motif õÂħÆ. ‧ 國. 學. щ5ŚȋE3 ʐɎşΘƀ ƒPINIT motif ͯRING finger motif ͎Ŋˋщ5ŚȋLǍ͂ ƓƆƿƺş ƷƓȆзPIAS3 şPINIT motif ̍ΎѠɽƵFş×̟(Duval et al., 2003). ‧. PIAS1 ;şRLD domain Ɠ NJʭícysteine şconserved domain |ȓͯõÂ. y. Nat. sit. RING finger motif şSiz / PIAS RING (SP-RING) Æч ʎNJ˓͕ͯщ5ŚȋE3 ʐɎş. er. io. λƏ(zinc finger) ˓͕ƆƨÜ ƒʎNJconserved domain ͯщ5ŚȋE3 ʐɎşƟŃÆ. al. iv n C Ʈѧ ̍ġɇşщ5ŚȋE3 h e nʐɎƟŃʨ PIAS gchi U n. Ɔ3şчŸ (Jackson, 2001) µÈ :ͥƓ¸ɽƵѫ ńŤƓѫF ƷƓɂPIAS1 ʅ +şRLD. ʅşC ͤÆȫƃş. serine / threonine ŶȁŃ PIASx ȴɎÂşPIASxα ıPIASxβ ƸC ͤȶȻ: Δ ĨοȶȻƧƓ Δş (Arora et al., 2003 Rytinki et al., 2009) È Ϲ̮7у͌. ¸C ͤȶȻʭíşŃȶȻacid domain (AD) ½¸SIM (SUMO1. Interaction Motif) ͯSUMO1̍ɰƨ>Þ(Shuai K., 2006) ÆεşƓ èġɂSIM {ȧĚ:̍ΎѠщ5ŚȋE3 ʐɎşƟŃ PIAS ʅBħÆщ5ŚȋE3 ʐɎşƟ Ń |˻Lщ5ŚȋͯΖȼµşÞ(Kotaja et al., 2002) o+]э˓ŏѩ PIAS ʅ ş°͢˓͕ͯщ5ŚȋLǍ͂şчŸ ΥƪĎ lƺƓɍȗʔ̼щ5ŚȋE3 ʐɎ PIAS1 ˬÔşщ5ŚȋLǍ͂Ɠé|β̝Ζʅδşмļ. 15.
(205) ɸG̝
(206) γ]<ƪĎşP˒ɱ ͌ѪƁ<źşƪĎ˓ŏѩ c-Jun N-terminal kinase 1 (JNK1|ġHes-1 Serine 263 ş×̟ˋЃL Ěˬ ćΊwHes-1 ʅδşмļoPΊɆHes-1 ЭϭĀĭ4͎ GluR1 ȼµşĀĭǩŏ(Lin et al., 2012) È|š ЭђƈǍ͂Þ͎ŊHes-1ƨ̚ƿ ƺ ˉÏ źHes-1 şЭђƈǍ͂ÞƨчƪĎЂ̭Ě:¹ µÈ Υγ]lеƓɍ ȗHes-1 Ɠé̍ˬÔ ͢ЭђƈǍ͂Þщ5ŚȋLǍ͂ oPǍ͂ƈ͎Hes-1 Ńδ ıxȓ+şΎѠ o*IÂʰЛĤɍȗɸ Л щ5ŚȋLǍ͂Ɠé|ΎѠHes-1 Ĩʅ δşмļ Зˉ<źƪĎʪϋщ5ŚȋLǍ͂ƺʔ̼E1 ƟLıE2 ˓³ʯȓò ÂΖʅşǍ͂Þ ÝƓ͎ŊɽƵF3¹ΒşʅδÏē щ5ŚȋLǍ͂şˋB ƺʔ̼E3 ʐɎşȷͯ ɸ!Л ЩE3 ʐɎ< ş PIAS1 oЂ̭Ɠé|ŷġ Hes-1 ʅδˬÔщ5ŚȋLǍ͂ DZώ͌ѪƁ<źşƪĎ˓ŏу͌PIAS1ȧ. þ͖щ. 政 治 大 al., 2011) ȧÈ< DZώ͌ѪƁ<źşƪĎÂŏ у͌ͯHes-1 ó¬ƨÜјŊ¯ NJʅ 立 δǛɘ<Hes-5 ͯǥљϊʁȖόƨч(Lee et al., 2007) ɸ)Л ɍȗʔ̼PIAS1 ˬÔ 5ŚȋE3 ʐɎşĒÒ -̍ЩǍ͂Эϭµ4ˬÏŷˬ3̈́şţ˱ϊʁȖό(Tai et. ‧ 國. 學. щ5ŚȋLǍ͂<ƈƓéHes-1 ʅδ̍ΎѠţ˱ϊʁȖόoPƓé̍β̝GluR1 ʅδ Ūɯ µÈ Υγ]lƺşƪĎΖϑɮ»*. ‧.
(207) ΡHes-1 Ɠéȓʔ̼щ5ŚȋE3 ʐɎPIAS1 ˬÔщ5ŚȋLǍ͂. y. Nat. !
(208) ̞̼щ5ŚȋǍ͂ƈşHes-1 Ɠé̍ΎѠĨʅδмļŃ. n. al. er. io. ɯ. sit. )
(209) ̞̼щ5ŚȋǍ͂ƈşHes-1 Ɠé̍ΎѠ3̈́ţ˱ϊʁͯȖόşŪɯPGluR1 şŪ. Ch. engchi. 16. i Un. v.
(210) 立. 政 治 大. Nat. y. ‧. ‧ 國. 學. $&. sit. n. al. er. io. +5 . Ch. engchi. 17. i Un. v.
(211) ɸ!ʜ͌ѪĆǪͯ^Ŗ ɸ ̝
(212) ɽƵȾξ(cell culture)
(213) ɽƵ̭Ǐ ͌ѪġşɽƵǴƓ"щƳƴ˙ѥɽƵhuman embryonic kidney 293T o*Уͣ HEK293T Ɉɣ͏˄ɟ;įvɽƵͥĬɽƵ̭ǏòÂş̼ˎƺ÷ʑˬÔ oАãµǬ˱ şŀŭÏʓÂUʖ¥ɽƵÉ/ §ɂíÆ10 % ƴhÓɤ(fetal bovine serum, FBS) ş Dulbecco's Modified Eagles Medium(DMEM, Life Technologies) ɽƵȾξɣ ̟Ŋ37 ℃ eǽΗ;̀ΝoÞƠ̖ʠ Ɉɣ͏˄ɟ;įvɽƵäǏͥƈ èɂɽƵͥʼn#37 ℃ eǽ ΗġͥF<ɽƵϞ̭ ƇĨ̭ǏòÂƈ į10 ml <ɽƵȾξɣ¶̒ɽƵͥFşɽƵ ¶̒ ̼ˎƺΪ͐5ZşʹɽƵ бZ1000 rpm
(214) 5 Iє Ɏ˝ §ìɏǓ§íÆ dimethyl. 政 治 大. sulfoxide (DMSO, Sigma) ùǏυşȾξɣĚw#10 ml ̋ЙşȾξɣ ªɂбZƈņõÂ. 立. şɽƵ͉̂Ϊ͐5ZşʹÂьǼɽƵɣ ϯ<ɂɽƵ͢ĬŠǤ10 HIɽƵȾξ; Ⱦ. ‧ 國. 學. ξ¸̕ƃ37 ℃
(215) 5 % CO2 <ϼÁȾξΤ; ̾RˬÔяrȾξ ̚яrȾξĬɸ)r Ě ˢĨɽƵŭŞŗмļƈ ^|ˬÔɽƵ͌Ѫ. ‧. !
(216) ɽƵȾξ. sit. y. Nat. HEK293T ɽƵ ȾξŊŠǤ10 HIɽƵȾξ; ɽƵȾξɣƠíÆ10 % ƴhÓ ɤ(fetal bovine serum, FBS) şDMEM Ⱦξɣ ɽƵȾξϿ͋Ơ̕ƃ37 ℃
(217) 5 % CO2 <. io. n. al. er. ϼ Á Ⱦ ξ Τ ; ¸ ˬ Ô ɽ Ƶ Ⱦ ξ ź § ɂ ɽ Ƶ Ⱦ ξ ɣ ̀ Ν Ĉ ) R ˬ Ô Ç я r. i Un. v. (subculture) IΠȾξ Dž§§ìɏȾξ;şȾξɣ ªw#UʉЃѰ̒ɣoɤƞʤο. Ch. engchi. şȾξɣ Ɏ˝ìɏЃѰ̒ɣĚw#η˭<̋ЙɽƵȾξɣ ɂǓ§ˤű¸Ⱦξ+<ɽ ƵΪ͐şʹ ʢƈįη˭<ɽƵьǼɣƿ̋w#ĬíÆ10 ml ɽƵȾξɣşŠǤ10 HI şȾξ; oͬƎĨŭ Ʒµ͌ѪˬÔ<ĉ ƺɂɽƵI˃Ĭ6 ń12 TΠ; Ż §ɂʹƈşɽƵ͉̂ьǼɣį20 µl ͯ20 µl ş0.4 % trypan blue (Invitrogen, USA) ɧ³ ïK ªɂɧ³ɣw¸ÓɭƻΒφ(Hemocytometer) +ǂƻΒɽƵ ʢƈo2.5x105 cell/ well ɂɽƵIΠĬTΠ; Ƈ̾_ɽƵŭŞŗмļƈè|ˬÔ͌Ѫ ̚ɽƵξÑư20 r ƿ̭̋ǏɽƵ )
(218) äǏɽƵ ¸ˬÔäǏɽƵź §̀ΝɽƵȾξɣ ɂŭÑ$I͚<ɽƵ §ìɏǓ§¸Ⱦξ ;şɽƵȾξɣ ϯ<w#̋ЙşȾξɣƈ ɂˤű¸Ⱦξ+<ɽƵ͉̂Ϊ͐ïKş ʹ Ã˴Ñ15 ml бZͥ бZ1000 rpm
(219) 5 Iє ƇĨбZòÂƈ §ìɏǓ§şɽƵ Ⱦξɣ įη˭<̋ЙɽƵȾξɣɂɽƵ͉̂ʹ ªͯ10 % DMSO (Sigma) ùǏυïK 18.
(220) ɧ³ ¸ĈNJäǏͥ;I̫1 ml ƇäǏ<ɽƵ Ɏ˝ ɂäǏͥʼn̟ĬɽƵäǏɁɵ4 F ̟Ŋ-80 ℃ «Τ24 5Ǭ ªɂ<ɷ#Ĭɣ͏˄ɟ;Ŷ½. ɸ!̝
(221) ɽƵЭƖ(transfection)
(222) ɽƵЭƖ ¸ ɽ Ƶ Э Ɩ ş ź R » + ƾ ņ ƾ § ̀ Ν ɽ Ƶ Ⱦ ξ ɣ ɂ ŭ Ñ $ I ͚ ş HEK293T ɽƵőЙɽƵȾξɣʹ ªoÓɭƻΒφ(Hemocytometer) ƻΒɽƵƈ o 2.5x105 cell/well <ɀƃɂɽƵIΠĬ6 ń12 TΠ ̟ʼnŊ̕ƃ37 ℃
(223) 5 % CO2 <ȾξΤ ; Ⱦ ξ ̾ _ è | ˬ Ô ɽ Ƶ Э Ɩ ͌ Ѫ ¸ ˬ Ô ɽ Ƶ Э Ɩ < ź § ̀ Ν ˈ Ó ɤ ş Dulbecco's Modified Eagles Medium (DMEM, Life Technologies) Ⱦξɣ γ]şɽƵЭ. 政 治 大 ˭бZͥ;w#100 µl ˈÓɤşDMEM Ⱦξɣ ¯Ǭ¸ NJ̫Æ100 µl ˈÓɤş 立 DMEMȾξɣş̅˭бZͥ; ğ̘Ĉµg <δѫϖƃį2.5 µl şLipofectamine 2000 ƖƓolipofection ş^ÁɂɡŪɯ<δѫЭɷț#ɽƵF Dž§ ¸̫ÆɡЭƖ<δѫş̅ TM. ‧ 國. 學. Invitrogen̯υˏѓ ɧ³ïKϰ̟ŊƁ̕Þ?Iєƈ ^|ɂĦŤɧ³ïKĚϰ̟ ŊƁ̕Þ20Iє ˬÔЭƖǬ ɂɧ³ɣΪ͐͗#TΠ; òÂƈɂTΠªʼn̟Ŋ̕ƃ. ‧. 37 ℃
(224) 5 % CO2 <ȾξΤ;Ⱦξ δѫЭƖƈȾξÑ24 5Ǭ |ªˬÔĨqƨч͌Ѫ». y. sit. io. n. al. er. ў͌ѪIő. Nat. пŝşʃɮƷ:ƺ̞̼Ĩqʃɮş͌Ѫ ŻȾξÑ48 5Ǭƈ è|ÃįɽƵoˬÔƈ. v. !
(225) ʅδïδɣş˛į ЭƖòÂĚj̞̼ ļǬ˱Þ<ɽƵ è|ÃįĨɽƵoˬÔƈў͌ѪIő Dž. Ch. engchi. i Un. § ìɏTΠFşɽƵȾξɣ oЃѰ̒ɣɤƞ Çƈ w#íÆʅδĀĭυ (protease inhibitor, Roche) PЃĀĭυ(phosphotase inhibitor, Roche) şïδΪέɣ (RIPA Lysis Buffer) oɂɽƵΫȆ ʼn̟¸ǂμЁφ(shaker) o120 rpm ̊ǭ'I є Ã˴ɽƵɣÑŊ1.5 ml <̅˭бZͥ Ŋ4 ℃
(226) 14000 rpm бZ'Iєo{ȧɽƵ˂ ϳ ÃįĨ+ɤɣèƠɽƵʅδïδɣ(cell lysate) Ŷ½Ŋ-80 ℃ «Τ. ɸ)̝
(227) Ö^ρЛŖ(Western blot)
(228) ʅδϖƃˇļPΔȣ͵ ͌ѪɒBradford^Ŗ ЩŃƖυProtein Assay Dye Reagent. (Coomassie. Brilliant Blue G-250Bio-Rad Labtories, USA) ˇļʅδïδɣ;ņíşʅδϖƃ Dž 19.
(229) § ɂProtein Assay Dye Reagent oͲжeˏѓ?dž ĈNJƇˇΔį2 µl w#1000 µl şProtein Assay Dye Reagent ˏѓɣ;ɧ³ïKƈ ̟ŊƁ̕QϹ2 Iє ªoI¦¦ƃ ƻ595 nm şŕŭȱˇĨì¦LJѧL è|ɇĬΔ;ʅδşϖƃ È^ŖşǓɮlƺƓ Coomassie Brilliant Blue G-250 ŃƖυͯʅδ˓³Ǭ ʢ3ì¦LJ̍Ɉ465 nm Эɷ Ĭ595 nm şǿŃ µÈЩI¦¦ƃƻ595 nm şŕŭȱˇʅδşϖƃ ĚoƴhÓɤ ʅ(bovine serum albumin, BSA) 2
(230) 4
(231) 6
(232) 8 μg/μl ʎ͢ϖƃÞƠΖ̖ϖƃÅΩ ɐͦ vȱˇ<Δşʅδϖƃ Δ̞̼ϖƃļ˭Iőƈ o5 džloading dye
(233) ̔˞ePΔ ʅδïδɣȣ͵ÂĈ1 µlíÆ1 µg şʅδ̒ɣ ɧ³ïKƈ ̟Ŋ95 ℃ wΝŐ (heat plate) wΝ 10 Iє èÂƠ|ƇIő<ʅδΔ ̞̼wΝ͵ʠºşʅδΔ| Ŷ½Ŋ-20 ℃ «Τ !
相關文件
Hague) 21, no.1 (Jan 1979) 1-19,以及”Two Problems in the History of Indian Buddhism: The Layman/Monk Distinction and the Doctrines of the Transference of Merit,” in
You are given the wavelength and total energy of a light pulse and asked to find the number of photons it
Students are asked to collect information (including materials from books, pamphlet from Environmental Protection Department...etc.) of the possible effects of pollution on our
Wang, Solving pseudomonotone variational inequalities and pseudocon- vex optimization problems using the projection neural network, IEEE Transactions on Neural Networks 17
volume suppressed mass: (TeV) 2 /M P ∼ 10 −4 eV → mm range can be experimentally tested for any number of extra dimensions - Light U(1) gauge bosons: no derivative couplings. =>
Define instead the imaginary.. potential, magnetic field, lattice…) Dirac-BdG Hamiltonian:. with small, and matrix
incapable to extract any quantities from QCD, nor to tackle the most interesting physics, namely, the spontaneously chiral symmetry breaking and the color confinement..
• Formation of massive primordial stars as origin of objects in the early universe. • Supernova explosions might be visible to the most
