title: A Minor Subset of Super Elongation Complexes Plays a Predominant Role in Reversing HIV-1 Latency.
abstract: Promoter-proximal pausing by RNA polymerase II (Pol II) is a key rate-limiting step in HIV-1 transcription and latency reversal. The viral Tat protein recruits human super elongation complexes (SECs) to paused Pol II to overcome this restriction. Despite the recent progress in understanding the functions of different subsets of SECs in controlling cellular and Tat-activated HIV transcription, little is known about the SEC subtypes that help reverse viral latency in CD4(+) T cells. Here, we used the CRISPR-Cas9 genome-editing tool to knock out the gene encoding the SEC subunit ELL2, AFF1, or AFF4 in Jurkat/2D10 cells, a well-characterized HIV-1 latency model. Depletion of these proteins drastically reduced spontaneous and drug-induced latency reversal by suppressing HIV-1 transcriptional elongation. Surprisingly, a low-abundance subset of SECs containing ELL2 and AFF1 was found to play a predominant role in cooperating with Tat to reverse latency. By increasing the cellular level/activity of these Tat-friendly SECs, we could potently activate latent HIV-1 without using any drugs. These results implicate the ELL2/AFF1-SECs as an important target in the future design of a combinatorial therapeutic approach to purge latent HIV-1.
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Zichong
authors.initials: Z
authors.lastname: Li
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA Innovation Center of Cell Signaling Network, School of Pharmaceutical Sciences, Xiamen University, Xiamen, Fujian, China.
authors.forename: Huasong
authors.initials: H
authors.lastname: Lu
authors.affiliation: Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, California, USA qzhou@berkeley.edu.
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zhou
mesh_terms: D000595:Amino Acid Sequence; D001483:Base Sequence; D064113:CRISPR-Cas Systems; D002460:Cell Line; D004247:DNA; D004268:DNA-Binding Proteins / Q000235:genetics; D055786:Gene Knockout Techniques; D015497:HIV-1 / Q000502:physiology*; D006801:Humans; D019169:Jurkat Cells; D008969:Molecular Sequence Data; D012097:Repressor Proteins / Q000235:genetics; D035602:Transcriptional Elongation Factors / Q000235:genetics / Q000502:physiology*; D017735:Virus Latency
chemical_list: C471348:AFF4 protein, human; D004268:DNA-Binding Proteins; C447613:ELL2 protein, human; D012097:Repressor Proteins; D035602:Transcriptional Elongation Factors; C081449:AFF1 protein, human; D004247:DNA
references.citation: Virology. 2014 Apr;454-455:328-39
references.pmid: 24565118
references.citation: Nat Med. 2014 Apr;20(4):425-9
references.pmid: 24658076
references.citation: Mol Cell. 2010 May 14;38(3):428-38
references.pmid: 20471948
references.citation: Mol Cell. 2010 May 14;38(3):439-51
references.pmid: 20471949
references.citation: J Virol. 2001 Dec;75(23):11336-43
references.pmid: 11689614
references.citation: EMBO J. 2003 Apr 15;22(8):1868-77
references.pmid: 12682019
references.citation: J Virol. 2004 Sep;78(17):9105-14
references.pmid: 15308706
references.citation: Proc Natl Acad Sci U S A. 1997 Apr 15;94(8):3639-43
references.pmid: 9108030
references.citation: FASEB J. 1998 Nov;12(14):1437-46
references.pmid: 9806752
references.citation: Mol Cell. 2005 Aug 19;19(4):535-45
references.pmid: 16109377
references.citation: EMBO J. 2006 Jan 11;25(1):139-49
references.pmid: 16319923
references.citation: Retrovirology. 2006;3:66
references.pmid: 17014716
references.citation: Proc Natl Acad Sci U S A. 2007 Aug 21;104(34):13690-5
references.pmid: 17690245
references.citation: J Virol. 2008 Dec;82(24):12291-303
references.pmid: 18829756
references.citation: Nat Rev Drug Discov. 2009 Jan;8(1):33-40
references.pmid: 19116625
references.citation: Mol Cell. 2010 Feb 12;37(3):429-37
references.pmid: 20159561
references.citation: J Virol. 2010 Jul;84(13):6425-37
references.pmid: 20410271
references.citation: Genes Dev. 2011 Jul 15;25(14):1486-98
references.pmid: 21764852
references.citation: Cell. 2011 Jul 8;146(1):92-104
references.pmid: 21729782
references.citation: Curr Opin HIV AIDS. 2011 Jan;6(1):4-11
references.pmid: 21242887
references.citation: J Mol Biol. 2011 Jul 29;410(5):896-916
references.pmid: 21763495
references.citation: Proc Natl Acad Sci U S A. 2011 Sep 6;108(36):E636-45
references.pmid: 21873227
references.citation: Cell Host Microbe. 2011 Nov 17;10(5):426-35
references.pmid: 22100159
references.citation: Cold Spring Harb Perspect Med. 2011 Sep;1(1):a007096
references.pmid: 22229121
references.citation: J Virol. 2012 Mar;86(6):3253-63
references.pmid: 22238306
references.citation: Mol Cell. 2012 May 11;46(3):325-34
references.pmid: 22483617
references.citation: Nat Rev Mol Cell Biol. 2012 Sep;13(9):543-7
references.pmid: 22895430
references.citation: Cell Rep. 2012 Oct 25;2(4):807-16
references.pmid: 23041316
references.citation: J Leukoc Biol. 2012 Dec;92(6):1147-54
references.pmid: 22802445
references.citation: Nucleic Acids Res. 2013 Jan 7;41(1):277-87
references.pmid: 23087374
references.citation: Proc Natl Acad Sci U S A. 2013 Jan 8;110(2):E123-31
references.pmid: 23251033
references.citation: Elife. 2013;2:e00327
references.pmid: 23471103
references.citation: Nat Protoc. 2013 Nov;8(11):2281-308
references.pmid: 24157548
references.citation: Science. 2014 Jan 3;343(6166):80-4
references.pmid: 24336569
references.citation: Proc Natl Acad Sci U S A. 2014 Jan 7;111(1):E15-24
references.pmid: 24367103
references.citation: Nucleic Acids Res. 2015 Jul 13;43(12):5868-79
references.pmid: 26007649