PMID- 24434558
OWN - NLM
STAT- MEDLINE
DCOM- 20140506
LR  - 20240210
IS  - 1091-6490 (Electronic)
IS  - 0027-8424 (Print)
IS  - 0027-8424 (Linking)
VI  - 111
IP  - 4
DP  - 2014 Jan 28
TI  - An RNAi screen for Aire cofactors reveals a role for Hnrnpl in polymerase release 
      and Aire-activated ectopic transcription.
PG  - 1491-6
LID - 10.1073/pnas.1323535111 [doi]
AB  - Aire induces the expression of a large set of autoantigen genes in the thymus, 
      driving immunological tolerance in maturing T cells. To determine the full 
      spectrum of molecular mechanisms underlying the Aire transactivation function, we 
      screened an AIRE-dependent gene-expression system with a genome-scale lentiviral 
      shRNA library, targeting factors associated with chromatin architecture/function, 
      transcription, and mRNA processing. Fifty-one functional allies were identified, 
      with a preponderance of factors that impact transcriptional elongation compared 
      with initiation, in particular members of the positive transcription elongation 
      factor b (P-TEFb) involved in the release of "paused" RNA polymerases (CCNT2 and 
      HEXIM1); mRNA processing and polyadenylation factors were also highlighted 
      (HNRNPL/F, SFRS1, SFRS3, and CLP1). Aire's functional allies were validated on 
      transfected and endogenous target genes, including the generation of lentigenic 
      knockdown (KD) mice. We uncovered the effect of the splicing factor Hnrnpl on 
      Aire-induced transcription. Transcripts sensitive to the P-TEFb inhibitor 
      flavopiridol were reduced by Hnrnpl knockdown in thymic epithelial cells, 
      independently of their dependence on Aire, therefore indicating a general effect 
      of Hnrnpl on RNA elongation. This conclusion was substantiated by demonstration 
      of HNRNPL interactions with P-TEFb components (CDK9, CCNT2, HEXIM1, and the small 
      7SK RNA). Aire-containing complexes include 7SK RNA, the latter interaction 
      disrupted by HNRNPL knockdown, suggesting that HNRNPL may partake in delivering 
      inactive P-TEFb to Aire. Thus, these results indicate that mRNA processing 
      factors cooperate with Aire to release stalled polymerases and to activate 
      ectopic expression of autoantigen genes in the thymus.
FAU - Giraud, Matthieu
AU  - Giraud M
AD  - Division of Immunology, Department of Microbiology and Immunobiology, Harvard 
      Medical School, Boston, MA 02115.
FAU - Jmari, Nada
AU  - Jmari N
FAU - Du, Lina
AU  - Du L
FAU - Carallis, Floriane
AU  - Carallis F
FAU - Nieland, Thomas J F
AU  - Nieland TJ
FAU - Perez-Campo, Flor M
AU  - Perez-Campo FM
FAU - Bensaude, Olivier
AU  - Bensaude O
FAU - Root, David E
AU  - Root DE
FAU - Hacohen, Nir
AU  - Hacohen N
FAU - Mathis, Diane
AU  - Mathis D
FAU - Benoist, Christophe
AU  - Benoist C
LA  - eng
GR  - U01AI074575/AI/NIAID NIH HHS/United States
GR  - R01 AI088204/AI/NIAID NIH HHS/United States
GR  - R37 DK060027/DK/NIDDK NIH HHS/United States
GR  - 12487/CRUK_/Cancer Research UK/United Kingdom
GR  - U01 AI074575/AI/NIAID NIH HHS/United States
GR  - R01 DK060027/DK/NIDDK NIH HHS/United States
GR  - DK060027/DK/NIDDK NIH HHS/United States
GR  - AI088204/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PT  - Research Support, Non-U.S. Gov't
PT  - Validation Study
DEP - 20140116
PL  - United States
TA  - Proc Natl Acad Sci U S A
JT  - Proceedings of the National Academy of Sciences of the United States of America
JID - 7505876
RN  - 0 (Heterogeneous-Nuclear Ribonucleoproteins)
RN  - 0 (Transcription Factors)
RN  - 0 (heterogeneous-nuclear ribonucleoprotein I, mouse)
SB  - IM
MH  - Animals
MH  - Cell Line
MH  - Gene Knockdown Techniques
MH  - Heterogeneous-Nuclear Ribonucleoproteins/genetics/*physiology
MH  - Humans
MH  - Mice
MH  - *RNA Interference
MH  - Transcription Factors/*genetics/physiology
MH  - Transcription, Genetic/*physiology
MH  - AIRE Protein
PMC - PMC3910647
OTO - NOTNLM
OT  - autoimmunity
OT  - hnRNP
OT  - negative selection
COIS- The authors declare no conflict of interest.
EDAT- 2014/01/18 06:00
MHDA- 2014/05/07 06:00
PMCR- 2014/07/28
CRDT- 2014/01/18 06:00
PHST- 2014/01/18 06:00 [entrez]
PHST- 2014/01/18 06:00 [pubmed]
PHST- 2014/05/07 06:00 [medline]
PHST- 2014/07/28 00:00 [pmc-release]
AID - 1323535111 [pii]
AID - 201323535 [pii]
AID - 10.1073/pnas.1323535111 [doi]
PST - ppublish
SO  - Proc Natl Acad Sci U S A. 2014 Jan 28;111(4):1491-6. doi: 
      10.1073/pnas.1323535111. Epub 2014 Jan 16.