THE 58,000-DALTON CELLULAR INHIBITOR OF THE INTERFERON-INDUCED DOUBLE-STRANDED RNA-ACTIVATED PROTEIN-KINASE (PKR) IS A MEMBER OF THE TETRATRICOPEPTIDE REPEAT FAMILY OF PROTEINS

被引:166
作者
LEE, TG [1 ]
TANG, N [1 ]
THOMPSON, S [1 ]
MILLER, J [1 ]
KATZE, MG [1 ]
机构
[1] UNIV WASHINGTON,SCH MED,DEPT MICROBIOL,SC-42,SEATTLE,WA 98195
关键词
D O I
10.1128/MCB.14.4.2331
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
PKR is a serine/threonine protein kinase induced by interferon treatment and activated by double-stranded RNAs. As a result of activation, PKR becomes autophosphorylated and catalyzes phosphorylation of the alpha subunit of protein synthesis eukaryotic initiation factor 2 (eIF-2). While studying the regulation of PKR in virus-infected cells, we found that a cellular 58-kDa protein (P58) was recruited by influenza virus to downregulate PKR and thus avoid the kinase's deleterious effects on viral protein synthesis and replication. We now report on the cloning, sequencing, expression, and structural analysis of the P58 PKR inhibitor, a 504-amino-acid hydrophilic protein. P58, expressed as a histidine fusion protein in Escherichia coli, blocked both the autophosphorylation of PKR and phosphorylation of the alpha subunit of eIF-2. Western blot (immunoblot) analysis showed that P58 is present not only in bovine cells but also in human, monkey, and mouse cells, suggesting the protein is highly conserved. Computer analysis revealed that P58 contains regions of homology to the DnaJ family of proteins and a much lesser degree of similarity to the PKR natural substrate, eIF-2alpha. Finally, P58 contains nine tandemly arranged 34-amino-acid repeats, demonstrating that the PKR inhibitor is a member of the tetratricopeptide repeat family of proteins, the only member identified thus far with a known biochemical function.
引用
收藏
页码:2331 / 2342
页数:12
相关论文
共 70 条
[1]  
AUSUBEL FM, 1989, CURRENT PROTOCOLS MO
[2]   TRANSLATIONAL REGULATION BY THE INTERFERON-INDUCED DOUBLE-STRANDED-RNA-ACTIVATED 68-KDA PROTEIN-KINASE [J].
BARBER, GN ;
WAMBACH, M ;
WONG, ML ;
DEVER, TE ;
HINNEBUSCH, AG ;
KATZE, MG .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1993, 90 (10) :4621-4625
[3]   FUNCTIONAL EXPRESSION AND CHARACTERIZATION OF THE INTERFERON-INDUCED DOUBLE-STRANDED-RNA ACTIVATED P68 PROTEIN-KINASE FROM ESCHERICHIA-COLI [J].
BARBER, GN ;
TOMITA, J ;
HOVANESSIAN, AG ;
MEURS, E ;
KATZE, MG .
BIOCHEMISTRY, 1991, 30 (42) :10356-10361
[4]   DETECTION OF PROTEIN-KINASE HOMOLOGS AND VIRAL RNA-BINDING DOMAINS UTILIZING POLYCLONAL ANTISERUM PREPARED AGAINST A BACULOVIRUS-EXPRESSED DS RNA-ACTIVATED 68,000-DA PROTEIN-KINASE [J].
BARBER, GN ;
TOMITA, J ;
GARFINKEL, MS ;
MEURS, E ;
HOVANESSIAN, A ;
KATZE, MG .
VIROLOGY, 1992, 191 (02) :670-679
[5]  
BARBER GN, IN PRESS P NATL ACAD
[6]   DEGRADATION OF THE INTERFERON-INDUCED 68,000-MR PROTEIN-KINASE BY POLIOVIRUS REQUIRES RNA [J].
BLACK, TL ;
BARBER, GN ;
KATZE, MG .
JOURNAL OF VIROLOGY, 1993, 67 (02) :791-800
[7]   MESSENGER-RNA DECAY - FINDING THE RIGHT TARGETS [J].
BRAWERMAN, G .
CELL, 1989, 57 (01) :9-10
[8]   HUMAN HOMOLOGS OF THE BACTERIAL HEAT-SHOCK PROTEIN DNAJ ARE PREFERENTIALLY EXPRESSED IN NEURONS [J].
CHEETHAM, ME ;
BRION, JP ;
ANDERTON, BH .
BIOCHEMICAL JOURNAL, 1992, 284 :469-476
[9]   CLONING OF THE CDNA OF THE HEME-REGULATED EUKARYOTIC INITIATION FACTOR-2-ALPHA (EIF-2-ALPHA) KINASE OF RABBIT RETICULOCYTES - HOMOLOGY TO YEAST GCN2 PROTEIN-KINASE AND HUMAN DOUBLE-STRANDED-RNA-DEPENDENT EIF-2-ALPHA KINASE [J].
CHEN, JJ ;
THROOP, MS ;
GEHRKE, L ;
KUO, I ;
PAL, JK ;
BRODSKY, M ;
LONDON, IM .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1991, 88 (17) :7729-7733
[10]   SINGLE-STEP METHOD OF RNA ISOLATION BY ACID GUANIDINIUM THIOCYANATE PHENOL CHLOROFORM EXTRACTION [J].
CHOMCZYNSKI, P ;
SACCHI, N .
ANALYTICAL BIOCHEMISTRY, 1987, 162 (01) :156-159