Isolation of a Caulobacter Gene Cluster Specifying Flagellum Production by Using Nonmotile Tn5 Insertion Mutants

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1982 Nov 1

PMID 16593248

Citations 16

Authors

M Purucker

R Bryan

K Amemiya

B Ely

L Shapiro

Affiliations

Soon will be listed here.

Abstract

Caulobacter crescentus assembles a single polar flagellum from protein components synthesized at a specific time in the cell cycle. Of the 26 genes required for flagellum production, at least 4 of them-flaY, E, F, and G-map together in a single cluster. We have isolated DNA from this region of the chromosome by using a nonmotile mutant with a Tn5 insertion into flaE. C. crescentus DNA carrying the Tn5-flaE region and adjacent sequences was cloned into pBR325 and selected by transposon-encoded kanamycin resistance. The resulting plasmid was used as a probe to isolate the flaE region from a wild-type gene bank and to determine the chromosomal location of several deletion and insertion mutations within the flaY/E/F/G cluster. At least three promotors and three major transcripts were shown to originate from the cloned gene cluster. The role of these genes in flagellar biogenesis was examined by immunoprecipitation of mutant cell extracts with antiflagellin antibody. Deletions extending rightward into this gene cluster eliminated one of the two flagellin proteins normally synthesized by C. crescentus. Mutations mapping to the left permitted synthesis of both normal flagellins but at significantly decreased levels. These results suggest that the leftward end of this cluster contains a region that may function in a regulatory capacity whereas the rightward end may contain sequences overlapping a flagellin structural gene.

Citing Articles

Methylation involved in chemotaxis is regulated during Caulobacter differentiation.

Shaw P, Gomes S, Sweeney K, Ely B, Shapiro L Proc Natl Acad Sci U S A. 1983; 80(17):5261-5.

PMID: 6577421 PMC: 384233.

Physical mapping and complementation analysis of transposon Tn5 mutations in Caulobacter crescentus: organization of transcriptional units in the hook gene cluster.

Ohta N, Swanson E, Ely B, Newton A J Bacteriol. 1984; 158(3):897-904.

PMID: 6327646 PMC: 215526. DOI: 10.1128/jb.158.3.897-904.1984.

Analysis of the pleiotropic regulation of flagellar and chemotaxis gene expression in Caulobacter crescentus by using plasmid complementation.

Bryan R, Purucker M, Gomes S, Alexander W, Shapiro L Proc Natl Acad Sci U S A. 1984; 81(5):1341-5.

PMID: 6324186 PMC: 344829. DOI: 10.1073/pnas.81.5.1341.

Generation of a Tn5 promoter probe and its use in the study of gene expression in Caulobacter crescentus.

Bellofatto V, Shapiro L, Hodgson D Proc Natl Acad Sci U S A. 1984; 81(4):1035-9.

PMID: 6322183 PMC: 344758. DOI: 10.1073/pnas.81.4.1035.

Organization and nucleotide sequence analysis of an rRNA and tRNA gene cluster from Caulobacter crescentus.

Feingold J, Bellofatto V, Shapiro L, Amemiya K J Bacteriol. 1985; 163(1):155-66.

PMID: 4008439 PMC: 219093. DOI: 10.1128/jb.163.1.155-166.1985.

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