ESCHERICHIA-COLI UMUDC MUTANTS - DNA-SEQUENCE ALTERATIONS AND UMUD CLEAVAGE

被引:32
作者
KOCH, WH
ENNIS, DG
LEVINE, AS
WOODGATE, R
机构
[1] NICHHD, VIRUSES & CELLULAR BIOL SECT, BETHESDA, MD 20892 USA
[2] US FDA, MOLEC BIOL BRANCH, WASHINGTON, DC 20204 USA
[3] NICHHD, MOLEC GENET LAB, BETHESDA, MD 20892 USA
来源
MOLECULAR AND GENERAL GENETICS | 1992年 / 233卷 / 03期
关键词
ESCHERICHIA-COLI UMUDC MUTANTS; SOS MUTAGENESIS; PCR AMPLIFICATION; DNA SEQUENCE ANALYSIS; CHEMILUMINESCENT IMMUNODETECTION;
D O I
10.1007/BF00265442
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
The products of the chromosomally encoded umuDC genes are directly required for mutagenesis in Escherichia coli. Strains with either umuD or umuC mutations are rendered phenotypically non-mutable. To ascertain the molecular basis of this non-mutability, we determined the DNA sequence alterations of seven chromosomal umuDC mutants. Six mutants (umuD1, umuD44, umuD77, umuC36, umuC25, and umuC104) were found to be single base-pair substitutions that resulted in missense mutations. The Tn5 transposon insertion mutation (umuC122) resulted in a missense mutation followed immediately by a termination codon, producing a truncated UmuC protein lacking 102 carboxyl-terminal amino acids. All of the mutations were found to reside in regions of the UmuD and UmuC proteins that share high homology with analogous proteins. Chemiluminescent immunoassays revealed that the umuD1, umuD44, and umuD77 mutations all resulted in a non-cleavable UmuD protein. Because UmuD cleavage is a prerequisite for mutagenesis, the lack of UmuD processing appears to be the molecular basis for the non-mutable phenotype in these strains. These studies re-emphasize the critical nature of the RecA-mediated cleavage of UmuD for inducible mutagenesis and provide insights into the functional domains of the UmuC protein.
引用
收藏
页码:443 / 448
页数:6
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