PMID- 31207241
OWN - NLM
STAT- MEDLINE
DCOM- 20200611
LR  - 20200611
IS  - 1089-8638 (Electronic)
IS  - 0022-2836 (Linking)
VI  - 431
IP  - 17
DP  - 2019 Aug 9
TI  - CRISPR/Cas9-based Knockout Strategy Elucidates Components Essential for Type 1 
      Interferon Signaling in Human HeLa Cells.
PG  - 3324-3338
LID - S0022-2836(19)30369-9 [pii]
LID - 10.1016/j.jmb.2019.06.007 [doi]
AB  - Type I interferons (IFNs) have a central role in innate and adaptive immunities, 
      proliferation, and cancer surveillance. How IFN binding to its specific receptor, 
      the IFN alpha and beta receptor (IFNAR), can drive such variety of processes is an open 
      question. Here, to systematically and thoroughly investigate the molecular 
      mechanism of IFN signaling, we used a CRISPR/Cas9-based approach in a human cell 
      line (HeLa) to generate knockouts (KOs) of the genes participating in the type 1 
      IFN signaling cascade. We show that both IFNAR chains (IFNAR1 and IFNAR2) are 
      absolutely required for any IFN-induced signaling. Deletion of either signal 
      transducer and activator of transcription 1 (STAT1) or STAT2 had only a partial 
      effect on IFN-induced antiviral activity or gene induction. However, the deletion 
      of both genes completely abrogated any IFN-induced activity. So did a double 
      STAT2-IFN regulatory factor 1 (IRF1) KO and, to a large extent, a STAT1 KO 
      together with IRF9 knockdown. KO of any of the STATs had no effect on the 
      phosphorylation of other STATs, indicating that they bound IFNAR independently. 
      STAT3 and STAT6 phosphorylations were fully induced by type 1 IFN in the 
      STAT1-STAT2 KO, but did not promote gene induction. Moreover, STAT3 KO did not 
      affect type 1 IFN-induced gene or protein expression. Type 1 IFN also did not 
      activate p38, AKT, or ERK kinase. We conclude that type 1 IFN-induced activities 
      in HeLa cells are mediated by STAT1/STAT2/IRF9, STAT1/STAT1, or STAT2/IRF9 
      complexes and do not require alternative pathways.
CI  - Copyright (c) 2019 Elsevier Ltd. All rights reserved.
FAU - Urin, Victoria
AU  - Urin V
AD  - Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, 
      Israel.
FAU - Shemesh, Maya
AU  - Shemesh M
AD  - Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, 
      Israel.
FAU - Schreiber, Gideon
AU  - Schreiber G
AD  - Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, 
      Israel. Electronic address: gideon.schreiber@weizmann.ac.il.
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20190615
PL  - Netherlands
TA  - J Mol Biol
JT  - Journal of molecular biology
JID - 2985088R
RN  - 0 (Antiviral Agents)
RN  - 0 (IFNAR1 protein, human)
RN  - 0 (IFNAR2 protein, human)
RN  - 0 (IRF1 protein, human)
RN  - 0 (IRF9 protein, human)
RN  - 0 (Interferon Regulatory Factor-1)
RN  - 0 (Interferon Type I)
RN  - 0 (Interferon-Stimulated Gene Factor 3, gamma Subunit)
RN  - 0 (STAT1 Transcription Factor)
RN  - 0 (STAT1 protein, human)
RN  - 0 (STAT2 Transcription Factor)
RN  - 0 (STAT3 Transcription Factor)
RN  - 0 (STAT3 protein, human)
RN  - 0 (STAT6 Transcription Factor)
RN  - 0 (STAT6 protein, human)
RN  - 156986-95-7 (Receptor, Interferon alpha-beta)
SB  - IM
MH  - Antiviral Agents/pharmacology
MH  - *CRISPR-Cas Systems
MH  - Gene Expression Regulation
MH  - Gene Knockout Techniques/*methods
MH  - HeLa Cells
MH  - Humans
MH  - Interferon Regulatory Factor-1/genetics
MH  - Interferon Type I/*genetics
MH  - Interferon-Stimulated Gene Factor 3, gamma Subunit
MH  - Phosphorylation
MH  - Receptor, Interferon alpha-beta/genetics
MH  - STAT1 Transcription Factor/*genetics
MH  - STAT2 Transcription Factor
MH  - STAT3 Transcription Factor/genetics
MH  - STAT6 Transcription Factor
MH  - Signal Transduction/*genetics
OTO - NOTNLM
OT  - antiviral activity
OT  - gene regulation
OT  - protein phosphorylation
OT  - signal transduction
OT  - type I interferon
EDAT- 2019/06/18 06:00
MHDA- 2020/06/12 06:00
CRDT- 2019/06/18 06:00
PHST- 2019/02/28 00:00 [received]
PHST- 2019/05/26 00:00 [revised]
PHST- 2019/06/06 00:00 [accepted]
PHST- 2019/06/18 06:00 [pubmed]
PHST- 2020/06/12 06:00 [medline]
PHST- 2019/06/18 06:00 [entrez]
AID - S0022-2836(19)30369-9 [pii]
AID - 10.1016/j.jmb.2019.06.007 [doi]
PST - ppublish
SO  - J Mol Biol. 2019 Aug 9;431(17):3324-3338. doi: 10.1016/j.jmb.2019.06.007. Epub 
      2019 Jun 15.