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PMID: 7568151 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Double-stranded-RNA-dependent protein kinase and TAR RNA-binding protein form homo- and heterodimers in vivo.

Proceedings of the National Academy of Sciences of the United States of America ·Vol. 92 ·No. 21 ·1995-10-10 ·Pages 9445-9

Cosentino GP, Venkatesan S, Serluca FC, Green SR, Mathews MB, Sonenberg N

Abstract

The yeast two-hybrid system and far-Western protein blot analysis were used to demonstrate dimerization of human double-stranded RNA (dsRNA)-dependent protein kinase (PKR) in vivo and in vitro. A catalytically inactive mutant of PKR with a single amino acid substitution (K296R) was found to dimerize in vivo, and a mutant with a deletion of the catalytic domain of PKR retained the ability to dimerize. In contrast, deletion of the two dsRNA-binding motifs in the N-terminal regulatory domain of PKR abolished dimerization. In vitro dimerization of the dsRNA-binding domain required the presence of dsRNA. These results suggest that the binding of dsRNA by PKR is necessary for dimerization. The mammalian dsRNA-binding protein TRBP, originally identified on the basis of its ability to bind the transactivation region (TAR) of human immunodeficiency virus RNA, also dimerized with itself and with PKR in the yeast assay. Taken together, these results suggest that complexes consisting of different combinations of dsRNA-binding proteins may exist in vivo. Such complexes could mediate differential effects on gene expression and control of cell growth.

MeSH Terms
Amino Acid Sequence Humans Molecular Sequence Data Protein Binding Protein Conformation Protein Serine-Threonine Kinases/genetics,metabolism RNA-Binding Proteins/genetics,metabolism Recombinant Fusion Proteins/metabolism Sequence Deletion Structure-Activity Relationship Transcriptional Activation eIF-2 Kinase
Chemicals
RNA-Binding Proteins Recombinant Fusion Proteins trans-activation responsive RNA-binding protein Protein Serine-Threonine Kinases eIF-2 Kinase
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Cosentino G P
Department of Biochemistry, McGill University, Montreal, QC Canada.
Venkatesan S
Serluca F C
Green S R
Mathews M B
Sonenberg N
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Article Info
Journal
Proceedings of the National Academy of Sciences of the United States of America
Abbr.
Proc Natl Acad Sci U S A
ISSN
0027-8424
DOI
10.1073/pnas.92.21.9445
Published
1995-10-10
Pages
9445-9
Language
English
Region
United States
NLM ID
7505876
PMCID
PMC40818
Subset
IM
Grants
NIAID NIH HHS · AI34552 · United States
External Links
PubMed source DOI: 10.1073/pnas.92.21.9445 PMC Full Text View journal details
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