Genetic Mapping of RpoD Implicates the Major Sigma Factor of Bacillus Subtilis RNA Polymerase in Sporulation Initiation

Overview

Journal Mol Gen Genet

Specialties Genetics
Molecular Biology

Date 1985 Jan 1

PMID 2997585

Citations 41

Authors

C W PRICE

R H Doi

Affiliations

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Abstract

We have mapped the chromosomal locus of rpoD, which encodes the major sigma factor of Bacillus subtilis RNA polymerase. The rpoD locus lay between aroD and lys, tightly linked to dnaE and inseparable from crsA. Marker order in this region was acf-aroD-dnaE-rpoD(crsA)-spoOG-lys. By transformation using cloned donor DNA from the rpoD region, we identified the gene immediately upstream of rpoD as dnaE, which coded for a 62,000 dalton protein essential for DNA replication. Both dnaE and rpoD were transcribed in the same direction, counterclockwise on the chromosome. The gene functions and organization in the rpoD region are thus similar to those of the E. coli sigma operon. We also used transformation to identify crsA47 as a mutation within the sigma coding region itself. The crsA alteration of sigma renders the sporulation process insensitive to glucose catabolite repression, and also restores sporulation ability to strains carrying early-blocked spoOE, spoOF, and spoOK mutations. Thus the major sigma factor and these spoO gene products directly or indirectly affect the same cellular function.

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References

Anagnostopoulos C, Spizizen J . REQUIREMENTS FOR TRANSFORMATION IN BACILLUS SUBTILIS. J Bacteriol. 1961; 81(5):741-6. PMC: 279084. DOI: 10.1128/jb.81.5.741-746.1961. View

Wiggs J, Gilman M, CHAMBERLIN M . Heterogeneity of RNA polymerase in Bacillus subtilis: evidence for an additional sigma factor in vegetative cells. Proc Natl Acad Sci U S A. 1981; 78(5):2762-6. PMC: 319437. DOI: 10.1073/pnas.78.5.2762. View

Laemmli U . Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970; 227(5259):680-5. DOI: 10.1038/227680a0. View

IONESCO H, Michel J, Cami B, Schaeffer P . Symposium on bacterial spores: II. Genetics of sporulation in Bacillus subtilis Marburg. J Appl Bacteriol. 1970; 33(1):13-24. DOI: 10.1111/j.1365-2672.1970.tb05230.x. View

de Bruijn F, Lupski J . The use of transposon Tn5 mutagenesis in the rapid generation of correlated physical and genetic maps of DNA segments cloned into multicopy plasmids--a review. Gene. 1984; 27(2):131-49. DOI: 10.1016/0378-1119(84)90135-5. View

Wang L, PRICE C, Doi R . Bacillus subtilis dnaE encodes a protein homologous to DNA primase of Escherichia coli. J Biol Chem. 1985; 260(6):3368-72. View

Travers A, Buckland R, Goman M, Le Grice S, Scaife J . A mutation affecting the sigma subunit of RNA polymerase changes transcriptional specificity. Nature. 1978; 273(5661):354-8. DOI: 10.1038/273354a0. View

Haldenwang W, Losick R . Novel RNA polymerase sigma factor from Bacillus subtilis. Proc Natl Acad Sci U S A. 1980; 77(12):7000-4. PMC: 350428. DOI: 10.1073/pnas.77.12.7000. View

Piggot P . Mapping of asporogenous mutations of Bacillus subtilis: a minimum estimate of the number of sporeulation operons. J Bacteriol. 1973; 114(3):1241-53. PMC: 285388. DOI: 10.1128/jb.114.3.1241-1253.1973. View

10.

Kawamura F, Doi R . Construction of a Bacillus subtilis double mutant deficient in extracellular alkaline and neutral proteases. J Bacteriol. 1984; 160(1):442-4. PMC: 214740. DOI: 10.1128/jb.160.1.442-444.1984. View

11.

Burton Z, Gross C, Watanabe K, Burgess R . The operon that encodes the sigma subunit of RNA polymerase also encodes ribosomal protein S21 and DNA primase in E. coli K12. Cell. 1983; 32(2):335-49. DOI: 10.1016/0092-8674(83)90453-1. View

12.

Wong S, PRICE C, Goldfarb D, Doi R . The subtilisin E gene of Bacillus subtilis is transcribed from a sigma 37 promoter in vivo. Proc Natl Acad Sci U S A. 1984; 81(4):1184-8. PMC: 344790. DOI: 10.1073/pnas.81.4.1184. View

13.

Kawamura F, Saito H . Isolation and mapping of a new suppressor mutation of an early sporulation gene spoOF mutation in Bacillus subtilis. Mol Gen Genet. 1983; 192(3):330-4. DOI: 10.1007/BF00392171. View

14.

Sanzey B . Modulation of gene expression by drugs affecting deoxyribonucleic acid gyrase. J Bacteriol. 1979; 138(1):40-7. PMC: 218235. DOI: 10.1128/jb.138.1.40-47.1979. View

15.

Lopez J, Freese E . Catabolite repression of enzyme synthesis does not prevent sporulation. J Bacteriol. 1980; 141(3):1447-9. PMC: 293851. DOI: 10.1128/jb.141.3.1447-1449.1980. View

16.

Soberon X, Covarrubias L, Bolivar F . Construction and characterization of new cloning vehicles. IV. Deletion derivatives of pBR322 and pBR325. Gene. 1980; 9(3-4):287-305. DOI: 10.1016/0378-1119(90)90328-o. View

17.

Sharrock R, Rubinstein S, Chan M, Leighton T . Intergenic suppression of spoO phenotypes by the Bacillus subtilis mutation rvtA. Mol Gen Genet. 2010; 194(1-2):260-4. DOI: 10.1007/BF00383525. View

18.

Sharrock R, Leighton T . Intergenic suppressors of temperature-sensitive sporulation in Bacillus subtilis are allele non-specific. Mol Gen Genet. 1981; 183(3):532-7. DOI: 10.1007/BF00268777. View

19.

Losick R, Pero J . Cascades of Sigma factors. Cell. 1981; 25(3):582-4. DOI: 10.1016/0092-8674(81)90164-1. View

20.

Horinouchi S, Weisblum B . Nucleotide sequence and functional map of pC194, a plasmid that specifies inducible chloramphenicol resistance. J Bacteriol. 1982; 150(2):815-25. PMC: 216434. DOI: 10.1128/jb.150.2.815-825.1982. View