ap- ˘ ˇˆ˙ˆ˝˛˚˜ ! transcriptional repression of human ... · transcriptional repression of...

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0 ECC 腞腜腑腒腓腊腔腍腋腏腚腙腢腨腣腡腣腚腙 0 ECC 腞腜腑腒腓腊腔腍腋腏腚腙腢腨腣腡腣腚腙 AP- . 腃腇腈 HIV 腐腕腌腋腔腎腄腧腄腦腠腤腛腘腝 Transcriptional Repression of Human Immunodeficiency Virus Type + by AP- . Kenichi IMAI 臇膣臘腷腃膭膡膜膭臃膹膧臐腭腧臗臮Department of Molecular and Cellular Biology, Nagoya City University Graduate School of Medical Sciences 腘腯腹腷膁膭膪臙 2 : 203+,**0 腅腀腆腃 腹腷膁腸腷膒膀腢腌腑腱腸腷膒膀膛腫腹腣臹膊膔腸腷膒膀腍腯腣臛腣膩腞腉腱腙腬腸腷膒膀腣 臯腎臡臝臛臫臊膵臋腢腏腐膘臼腖腱臛腣膩腞腤膊膔腸腷膒膀 DNA 臍臭腣臑腢腼膇膍 RNA RNase H 腢腮腛腝膫腓腲腱腙腬膛腫腹腣臹腤腌腒腯腡 腒腲腫腞腣 HIV 臛臃膹腍腯腢臵膰致臐腍腯 腣腸腷膒膀臯腣膯臬膤腤臡臝臐臛膯臬膤膠臗 NF-kB 腟腸腷膒膀腣臚腜膅膐膕膀膯臬膤膠臗 Tat 腢腮腛 腢臫臊腓腲腝腊腱腒腟腎腯腍腟腡腛腙 +NF-kB 腵腤腕腬腟腖腱 DNA 膿臍臬臛膯臬膤膠臗腣 LTR 腨腣膿臍腤腽腶腻膄膋腃膃腃腣臓腢腮腱膈膀膅 膕腶膂膄膒膤腢 腋腻膔膌膄膕臋臺腣 膱腟 TBP TFIID腣膳腯腑臛膠臗腭 RNA polymerase II 腣膑腻 膒腃膅膎膕膅腵臻腖腀腬Tat 臛腣臖腫腛腙腥腍腰 mRNA TAR 膞腢腢膿臍腔臛臦臻臧 膠臗腞腉腱 P-TEFb 腵膑腻膒腃膅腖腱腒腟腢腮腰 HIV 膔腸腷膒膀腣臛腵腔腐臤臢腓腗腱HIV 臛膯臬膤膵臋腎臛膠臗膓膋膒腉腱腊腤腻 膔膌膄膕膓膋膒腞腯腍腟腡腛腙膟HIV 臛腣膎腺膆膁膍腢膲腔腝腤腠腱HIV 臆腣臫膵 臋腎臛膓膋膒腞腯腍腢腡腲腥膰致膚臚膵臋腣膫 腨腟腜腡腎腱腒腟腎膴腞腏腱腒腲腫腞腢臛膠臗 YY-+ HDAC LTR 腢膑腻膒腃膅腖腱腒腟腢腮腰膀膅膕腣腶膂膄膒膤腵腦腏腌腒腔腻膔膌膄膕膓膋膒腞 HIV 臛腵臫腔腝腊腱腒腟腎臎腓腲腝腊腱 ,腫腙LTR 臤腣 negative regulatory element NRE腢膿臍腖腱臛膠臗腎 HIV 臛腵臫腖腱腟腊腋臎腎腉腱腎腔腊臗膵臋腤腴腍腛腝腊腡腊 -臜腯腤HIV LTR TATA box 臛膠臗 AP- . 腣膿臍腾腷膅腎臼臒腔AP-. HIV 臆腣臫膠臗腟 腔腝膵腔腝腊腱腒腟腵臅腊腚腔腙 .腦腠腖腟 AP-. HIV LTR 膗臶臜腯腤Tat 臫臊膛臗腵腢臂臔腔腙膿 DNA 臌臷腞腉腱 OGG+ 腕腎腔腙Tat 腢腮腱 臣臐腡 OGG+ 臆腣腳腖膵臋腵膫臰腔腙腟腒腳AP-. OGG+ 臆腵腢臫臊腔腝腌腰Tat AP-. 腟膿臍腔腝腘腣臓臫腖腱腒腟腞 OGG+ 臛腵腔腝腊腱腒腟腵臅腊腚腔腙 /腘腣臉腣膫臰腞HIV LTR TATA box 腅腇,,腄腇+1AP-. 腣膿臍腾腷 膅腎臼臒腖腱腒腟腓腯腢臞HIV 腾膉膃腷膊 LTR AP-. 膿臍腾腷膅腵腩腙膿膨腾膉膃腷膊 BAC 腔腁 腐腣腾膉膃腷膊腞 AP-. 腣膿臍腛腻 CAGCTG臼腓腲腝腊腱腒腟腢腉腲腔腙AP-. +/ 腙腏臶腢 Tjian 腯腢腮腛腝 SV.* late gene 腵膯臬膤腖腱膠臗腟腔腝臅腜 腍腛腙 HLH-Zip 膽腣臛膠臗腞腉腱 0腘腣臉腣臃膹腞TGF-b Caspase 3, Angiotensinogen 腡腠腣膛臗膊膔 膏腃膃腃腢 AP-. 腣膿臍腾腷膅腎臼臒腖腱腒腟腎臎腓腲 腙腎臸臈臓膠臗腵腤腕腬 AP-. 腢腮腱臣臐腡膛臆臫臊膵臋腢腜腊腝腤膟臱臎腎腡腊AP-. 膿臍腾腷膅 LTR TATA box 腢臼臒腔腫腙腘腣腛腻臼腓腲腝腊腱腒腟腍腯AP-. HIV 臛腢腌腊腝膥腯 腍腣臓腵膷腪腔腝腊腱膦臬腎臨臕腓腲腙AP-. HIV 腄腦腠腉腧腃腤腛腁腈 腤腕腬腢AP-. HIV 臛腢膷腪腖膢膺腵 T 腮腧膸膾臐腊腝 Luciferase assay 腢腝腩腙腼腶: 臏膝臁膟 腅腈.01**-+ 臇膣臘臩膼臩至自 + 臇膣臘腷腃膭膡膜膭臃膹膧臐腭腧 臗臮膭腆 Fax : */,2/3+,-/ ,**0 / +3 ,**0 The Japanese Society for AIDS Research The Journal of AIDS Research ,* 20

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Page 1: AP- ˘ ˇˆ˙ˆ˝˛˚˜ ! Transcriptional Repression of Human ... · Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-. Kenichi IMAI ... LTR ^ ]

� 0� ECC�������� ������ �� 0� ECC�������� ������ �

AP-.���HIV������������ !Transcriptional Repression of Human Immunodeficiency

Virus Type + by AP-.

� � � �Kenichi IMAI

������������ �������Department of Molecular and Cellular Biology,

Nagoya City University Graduate School of Medical Sciences

�������� 2 : 20�3+� ,**0

" # $ �

������������������ !�"�#� $%����&' ( "�)*�+,���� �-./0 (1234��56789�: ; ( "�)#$%����DNA<= >�?@A RNA. RNase

H�BCD�EFG�+,��� !�#�H'IJ: HGK) HIV (��&'���L�MN��&' �����- OPQ#� /0�� (OPQR�NF-kBS���� TUVWX�OPQR� Tat�BCD�Y��12FGDJ�HS.�'&SIC++Z:NF-kB[#\,S9�DNA]<P (OPQR� LTR^ ]<#� _`abcded f��B�g�VX`hb�Q��ia%jbX4k �lS TBP

mTFIIDZ � n� (R�o RNA polymerase II pa�dVqXV[r9: ��� Tat# ( sKC+t&u mRNA TAR�v��w��]<x� (y�rzR�)*� P-TEFb[pa�dV9�HS�BuHIV$%���� ([�x6{|F}�:

HIV (OPQ34.� (R�~c�� *�J#a%jbX~c�)�'&SIC+��)� HIV ( � q���A��xD#��I�. J: HIV!� "134. (~c�)�'&�IGt� L�MN�T34 E�^SUI.�HS.�#)5�: HGK)�� (R�YY-+.HDAC[ LTR�pa�dV9�HS�Bu� g�VX $`hb�Q[�5�Hxa%jbX~c�)HIV (["1xDJ�HS.�FGDJ�,Z: K+�

LTR{ negative regulatory element mNREZ �]<9� (R�.HIV (["19�SJi�.*�.� �xJ��34#�&CDJIJ-Z:%�'#� HIV LTR TATA box�&� (R� AP-

. ]<��V.8�x� AP-..HIV!� "1R�SxD3'xDJ�HS[�J�x+.Z:

��%& AP-.'HIV LTR

��%�'#� Tat(12��[)�����x+]�� DNA�*��)*� OGG+[+,x+: Tat�B���I OGG+��!� -.34[E�x+SH��AP-.. OGG+ !�[��12xD�u� Tat# AP-.

S]<xD� f�["19�HS) OGG+ ([-.xDJ�HS[�J�x+/Z: � � E�)� HIV

LTR TATA box�& m�,,��+1Z � AP-. ]<��V.8�9�HS� F'��� HIV� e�$ LTR AP-.]<��V[/¡+]�� � e�$ B� A� C� 6 � e�$) AP-. ]<01 mCAGCTGZ .2638FGDJ�HS�45x+: AP-.# +/6��� Tjian

'�BCD SV.* late gene[OPQ9�R�SxD�U&C+HLH-Zip¢ (R�)*�0Z: � � ��)�TGF-bo Caspase 3, AngiotensinogenI£ ��$%¤ded� AP-. ]<��V.8�9�HS.�FG+.� ¥¦f�R�[#\, AP-.�B���I��!�1234�UJD#�§�.IJ: AP-.]<��V. LTR TATA box�&�8�x� K+� 01.2638FGDJ�HS&'� AP-..HIV (��JD¨'& f�[©ªxDJ�«'P.¬­FG+:

AP-."HIV���(����)�

#\,�� AP-..HIV (�©ª9®¯[ T���B°7±²��[�JD Luciferase assay�D/¡+: �

��89³ : ���� m´.01�**-+ µ:¶����·;¸·;<¹º + ������������ �������ZFax : */,�2/3�+,-/

,**06 /» +3�¼=

�,**0 The Japanese Society for AIDS Research The Journal of AIDS Research

m ,* Z20

Page 2: AP- ˘ ˇˆ˙ˆ˝˛˚˜ ! Transcriptional Repression of Human ... · Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-. Kenichi IMAI ... LTR ^ ]

���� AP-.� basal������ Tat TNF-a���HIV����������� �� +A�� LTR�AP-.�� �!"#$�!"% LTR�&'(���)*�+,-./01�� 23�45�� AP-.6��78��(9:�('�0;0�<=��,�� AP-.�>?� siRNA�@A�BC����� HIV�����D�EF���� +B��GE���0-� AP-.�HIV����78��(9:�('�H�6I-0�/1��

AP-.�HLH-Zip%��78JK� NLM�DNA

�N�OP�HLHQR�S� dimerization�OP� leu-

cine repeat (LR)� CLM�� Q,P-rich� acidicQR�S6T5?�1�� HIV����&U� AP-.�9:QR�S

�<=��,�� VW�!"% AP-.�)X�Y�Z��[\��� Y���� HLHQR�S�]'�!"% AP-.

���Z�6+,-./01�H�0-� HIV������DNA���N6^_J�H�6P01��

AP-.���HIV�����

AP-.6`��a/RbcdeHIV������('��0fg�h,�� AP-.��N�� 6 TATA box

�ij�T5?�H�0-� AP-.6 TBP� TATA boxk��N�lm�('�n:�6op#.�� H�q� AP-

.�rstuvw�)X� EMSA�([\����� AP-

.� TBP� TATA boxk��N�xyzT{�����

� + AP-.� HIV���|��}?�A. CEM@A� LTR�BC� Lucirerase assay�~1���� AP-.� HIV�������� B.

,3-@A� siRNA�BC�45�� AP-.���������S#$�� ����� HIV����6EF�� �����

The Journal of AIDS Research Vol. 2 No. , ,**0

� ,+ �21

Page 3: AP- ˘ ˇˆ˙ˆ˝˛˚˜ ! Transcriptional Repression of Human ... · Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-. Kenichi IMAI ... LTR ^ ]

�� ,A�� AP-.� TBP���� in vivo� �������� AP-.���HIV������ TBP������� �!"#$% �� ,B��

AP-.���HIV���&�' TATA box�()*+,�-�./0���121345�%6�� LTR7� AP-

.�89:;<3 TATA box1=>0� AP-.�?@3AB0%�C�%6��DE� AP-.89:;<3FG#H%LTR�� TATA box1=IJ�K>�L�M%� AP-.:;<3NO0%FGP LTR3Q0 Luciferase assay3R %�C�8@�STP LTR�� ������ AP-.:;<3 TATA box1=>$%UV�NO0% LTR� �

�� AP-.�����?@'�6=$%� W7�8@1=�AP-.� TBP�()*+,WX��� Y=1�Z[\]^�HIV���3&�0���_�'`a#$%�bc� ���&�de� ��HDAC���fg(h

+ijk��&�'lm#$���� C_�� AP-.�HIV��&�� n�HDAC�op3AB0%�AP-.�HDAC'89q�rst3`a0uvwxy��45%8@� AP-.�z{t�HDAC+� �| ,�89q�_�'�6=$% �� -A�� }~�� AP-.���&��HDAC'op0���1213��q�%6��HDAC������� TSA3��AB0%8@� TSA

� , AP-.� TBP� TATA box��893��q�A. HIV LTR3�% EMSA assay. AP-.���./�� TBP� TATA box��893��0%�B. Lucirerase assay. TBP������ AP-.��� HIV�����?@'!"#$%�

� - AP-.� HDAC�89q�A. AP-.���uvwx3R %8@� AP-.�z{t� HDAC+� ,�890%� B. HDAC������� TSA����� AP-.��� HIV�����?@'!"#$%�

K Imai : Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-.

� ,, �22

Page 4: AP- ˘ ˇˆ˙ˆ˝˛˚˜ ! Transcriptional Repression of Human ... · Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-. Kenichi IMAI ... LTR ^ ]

����� AP-.���HIV���� ������ �� -B��

HIV������ LTR�� AP-.�� ����

���HIV������� AP-. TBP! HDAC�"#$%&��'�! HIV()*+�����,� ACH, U+��$ -./012345678$9-:;<��=�>?! @ABC#����D-.� LTRE�FGHI AP-. HDAC+�>JKL'M�! TNF-a���$ABN� ! LTROM AP-. HDAC+KPQ<! R� TBP

pol IIK LTR�S/TUV��.W�X ! �M�YZV3�[\2T]KDX^.-�X KL'M�� ��.A�� AP-.�>J_`V$ab�c� LTR$���de</01234567$9^�>?! S/TUV���HDAC+Kfg<�X OM! AP-.KHDAC+$ LTR�S/TUV<.-�X Khi��� �� .B��

HIV������ AP-.����

HIVjk�lN� AP-.�m?$:;N��'�!Jurkat T��� pNL.--$de< p,. ELISA assay$9^�� =�>?! AP-.� basalnoT��p`TZjk TNF-a���HIV�jk$qr<� �� /A�� st! AP-

. siRNA$ pNL.-- u����deN� HIV�jk�R�vw<� �� /B��xv�>?OM! AP-.K TATA box�1Zy3z

HDAC�S/TUV��^.HIV��${�r|<.-�X K}MO I^�� HIV()*+�����D-.! @AB�C#�� AP-.K TATA box$1Zy3zN� ~��HDAC$ LTR�S/TUV<.-�X !TNF-a�ABKe� AP-. HDACK LTROMPQ<���� TBP pol IIK LTR�S/TUV���>?$��J�c� ! ������� AP-.�"#KHIV�*+�������I��$�^.-���)Khi����

����

�����������������HIV�K AIDS

���D�����)p`TZ����� �W ¡N�X K}MO�I^�2�� X�X OM! HIVjk$qrN�X � AIDS��K¢£<! HAART�8�¤m)K¥vN�X K¦§���� <O<! ¨©p`TZ�K:�ª«x¬�I^.uID*+����K­®N�X K¯° I^.-�� ±^.! AIDS��$²-³'���*+�����´0p`TZOM��µ)]$³N�X K¶·�,� ��M��� ¸t�! *+����OM�p`TZjk$��DX<! HAART ¹ºJ�c�

� . HIV()*+�����D»� ChIP assay

A. ACH,! UI��$ -. ChIP assay$9^�>?! @ABC#����D-.� LTRE�FGHI AP-. HDAC+�>JKL'M��� TNF-aAB¼�� AP-. HDAC+KPQ<! R� TBP

pol IIK LTR�S/TUV��.W�X ! �M�YZV3�[\2T]KDX^.-�X KL'M��� B. AP-.�>J_`V$ab�c� LTR$ -� ChIP assay��! S/TUV���HDAC+Kfg<��

The Journal of AIDS Research Vol. 2 No. , ,**0

� ,- �23

Page 5: AP- ˘ ˇˆ˙ˆ˝˛˚˜ ! Transcriptional Repression of Human ... · Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-. Kenichi IMAI ... LTR ^ ]

������������� ������������������ ������HIV� T��� HDAC+ !" valproic acid 3#$ IL-1 +*#���%&'( �)�*+,-������.�/0 %����1 ���� 2 3�4( HAART5���6789:"*;<=�3>*?@A*BC�D53� EFGHIJKL# ��24( �*��3��5=%�M�NO *789:��/4����5P�-( 2M����*HIV

�Q$�R�S���%T( ��U*�V'�W���X��������

AIDS�=HAART���YZ%3�[�\M��]�%�( HIV=�^*_����� `a�Sb�cd%T( ef*��� AIDS�g���� �-��U�V5������h=3<( i%�j�$�"kS789:*le3>;<*m��n���oY( pqr*AIDS����g���*s��t��3����� �43�HIV*�u vwx*y!=( Z%3 AIDS��*s��d3������z5P��

�� : �*"= ECC{|}~LK9�8����#��������%�P�Y������i %� $�*���*��%(&����'���%�$����=�T������%�o(� ��i��

� �

+ # Price DH : P-TEFb, a cyclin-dependent kinase control-

ling elongation by RNA polymerase II. Mol Cell Biol ,* :

� / HIV)����� AP-.**�V'A. Jurkat T�� pNL.--�'+ ELISA�� p,.���, %� B. ,3-�� AP-. siRNA�'+ %&'( HIV*)���� %�

� 0 AP-.��� HIV�e**�}� �¡-¢£¤¥*�5=( NF-kB p/*( YY-+� HDAC

� LTR�LH9¦§������1������AP-.= TBP*��� HDAC*LH9¦§��Y¨.� HIV*�e�/��© ����ªM4��� TNF-a3>*¢£�+�� NF-kB0*«S¬�1�LH9¦§���<�­KHI®¦¯¦3>*j���Y( H°±IG*²³�o�Y LTR24 AP-.� HDAC�23��� ´*µ( TBP3>*¶$�u·¸¹�º�»i� HIV*�u�¼i��ªM4���

K Imai : Transcriptional Repression of Human Immunodeficiency Virus Type + by AP-.

E ,. #3*

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integrated promoter of human immunodeficiency virus

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HIV-+ Tat. J Biol Chem ,2* : ,01*+�,01+-, ,**/.

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