MOLECULAR-CLONING, SEQUENCING, AND OVEREXPRESSION OF THE STRUCTURAL GENE ENCODING THE DELTA-SUBUNIT OF ESCHERICHIA-COLI DNA POLYMERASE-III HOLOENZYME

被引:18
作者
CARTER, JR
FRANDEN, MA
AEBERSOLD, R
MCHENRY, CS
机构
[1] UNIV COLORADO,HLTH SCI CTR,DEPT BIOCHEM BIOPHYS & GENET,4200 E 9TH AVE,DENVER,CO 80262
[2] UNIV BRITISH COLUMBIA,BIOMED RES CTR,VANCOUVER V6T 1Z3,BC,CANADA
[3] UNIV BRITISH COLUMBIA,DEPT BIOCHEM,VANCOUVER V6T 1Z3,BC,CANADA
关键词
D O I
10.1128/JB.174.21.7013-7025.1992
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Using an oligonucleotide hybridization probe, we have mapped the structural gene for the delta subunit of Escherichia coli DNA polymerase III holoenzyme to 14.6 centisomes of the chromosome. This gene, designated holA, was cloned and sequenced. The sequence of holA matches precisely four amino acid sequences obtained for the amino terminus of delta and three internal tryptic peptides. A holA-overproducing plasmid that directs the expression of delta up to 4% of the soluble protein was constructed. Sequence analysis of holA revealed a 1,029-bp open reading frame that encodes a protein with a predicted molecular mass of 38,703 Da. holA may reside downstream of rlpB in an operon, perhaps representing yet another link between structural genes for the DNA polymerase III holoenzyme and proteins involved in membrane biogenesis. These and other features are discussed in terms of genetic regulation of delta-subunit synthesis.
引用
收藏
页码:7013 / 7025
页数:13
相关论文
共 81 条
[1]   INTERNAL AMINO-ACID SEQUENCE-ANALYSIS OF PROTEINS SEPARATED BY ONE-DIMENSIONAL OR TWO-DIMENSIONAL GEL-ELECTROPHORESIS AFTER INSITU PROTEASE DIGESTION ON NITROCELLULOSE [J].
AEBERSOLD, RH ;
LEAVITT, J ;
SAAVEDRA, RA ;
HOOD, LE ;
KENT, SBH .
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA, 1987, 84 (20) :6970-6974
[2]  
BIRNBOIM HC, 1979, NUCLEIC ACIDS RES, V7, P1513
[3]   PROGRAMMED RIBOSOMAL FRAMESHIFTING GENERATES THE ESCHERICHIA-COLI DNA POLYMERASE-III GAMMA SUBUNIT FROM WITHIN THE GAMMA-SUBUNIT READING FRAME [J].
BLINKOWA, AL ;
WALKER, JR .
NUCLEIC ACIDS RESEARCH, 1990, 18 (07) :1725-1729
[4]   A COMPLEMENTATION ANALYSIS OF RESTRICTION AND MODIFICATION OF DNA IN ESCHERICHIA COLI [J].
BOYER, HW ;
ROULLAND.D .
JOURNAL OF MOLECULAR BIOLOGY, 1969, 41 (03) :459-&
[5]   THE OPERON THAT ENCODES THE SIGMA-SUBUNIT OF RNA-POLYMERASE ALSO ENCODES RIBOSOMAL PROTEIN-S21 AND DNA PRIMASE IN ESCHERICHIA-COLI-K12 [J].
BURTON, ZF ;
GROSS, CA ;
WATANABE, KK ;
BURGESS, RR .
CELL, 1983, 32 (02) :335-349
[6]  
CARTER JG, UNPUB
[7]  
Chou P Y, 1978, Adv Enzymol Relat Areas Mol Biol, V47, P45
[8]  
DEBOER HA, 1986, MAXIMIZING GENE EXPR, P225
[9]  
Dong Z., UNPUB
[10]  
FAY PJ, 1981, J BIOL CHEM, V256, P976