The Haemophilus Influenzae DprABC Genes Constitute a Competence-inducible Operon That Requires the Product of the TfoX (sxy) Gene for Transcriptional Activation

Overview

Journal J Bacteriol

Publisher American Society for Microbiology

Specialty Microbiology

Date 1997 Aug 1

PMID 9244270

Citations 21

Authors

S Karudapuram

G J Barcak

Affiliations

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Abstract

We previously showed that dprA is required for efficient processing of linear DNA during cellular transformation in Haemophilus influenzae. In this study the transcriptional regulation of dprA and two downstream genes, dprB and dprC, is examined. We demonstrate by Northern blot analysis that the dprABC genes are transcriptionally coregulated and competence inducible. We used primer extension analysis to map the transcriptional start site of dprA and of rec-2, another transformation gene involved in DNA processing. Based upon these results, we were able to identify a 26-bp dyad symmetry element immediately upstream of the -35 regions of the predicted promoters of dprA, rec-2, and two other transformation genes, comA and pilA. Finally, using transcriptional fusions of dprA to the Escherichia coli lacZ gene, we show that expression of dprA::lacZ requires tfoX and that the presence of multiple copies of tfoX abolishes the temporal regulation of dprA, resulting in its constitutive expression.

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References

HERRIOTT R, Meyer E, Vogt M . Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol. 1970; 101(2):517-24. PMC: 284936. DOI: 10.1128/jb.101.2.517-524.1970. View

Gwinn M, Yi D, Smith H, Tomb J . Role of the two-component signal transduction and the phosphoenolpyruvate: carbohydrate phosphotransferase systems in competence development of Haemophilus influenzae Rd. J Bacteriol. 1996; 178(21):6366-8. PMC: 178514. DOI: 10.1128/jb.178.21.6366-6368.1996. View

WILCOX K, Smith H . Isolation and characterization of mutants of Haemophilus influenzae deficient in an adenosine 5'-triphosphate-dependent deoxyribonuclease activity. J Bacteriol. 1975; 122(2):443-53. PMC: 246077. DOI: 10.1128/jb.122.2.443-453.1975. View

Feinberg A, Vogelstein B . A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983; 132(1):6-13. DOI: 10.1016/0003-2697(83)90418-9. View

Martinez E, Bartolome B, de la Cruz F . pACYC184-derived cloning vectors containing the multiple cloning site and lacZ alpha reporter gene of pUC8/9 and pUC18/19 plasmids. Gene. 1988; 68(1):159-62. DOI: 10.1016/0378-1119(88)90608-7. View

Barcak G, Tomb J, Laufer C, Smith H . Two Haemophilus influenzae Rd genes that complement the recA-like mutation rec-1. J Bacteriol. 1989; 171(5):2451-7. PMC: 209920. DOI: 10.1128/jb.171.5.2451-2457.1989. View

Tomb J, Barcak G, Chandler M, Redfield R, Smith H . Transposon mutagenesis, characterization, and cloning of transformation genes of Haemophilus influenzae Rd. J Bacteriol. 1989; 171(7):3796-802. PMC: 210127. DOI: 10.1128/jb.171.7.3796-3802.1989. View

Langermann S, Wright A . Molecular analysis of the Haemophilus influenzae type b pilin gene. Mol Microbiol. 1990; 4(2):221-30. DOI: 10.1111/j.1365-2958.1990.tb00589.x. View

Coppola G, Huang F, Riley J, Cox J, Hantzopoulos P, Zhou L . Sequence and transcriptional activity of the Escherichia coli K-12 chromosome region between rrnC and ilvGMEDA. Gene. 1991; 97(1):21-7. DOI: 10.1016/0378-1119(91)90005-v. View

10.

ONeill M . Training back-propagation neural networks to define and detect DNA-binding sites. Nucleic Acids Res. 1991; 19(2):313-8. PMC: 333596. DOI: 10.1093/nar/19.2.313. View

11.

Larson T, GOODGAL S . Sequence and transcriptional regulation of com101A, a locus required for genetic transformation in Haemophilus influenzae. J Bacteriol. 1991; 173(15):4683-91. PMC: 208145. DOI: 10.1128/jb.173.15.4683-4691.1991. View

12.

Redfield R . sxy-1, a Haemophilus influenzae mutation causing greatly enhanced spontaneous competence. J Bacteriol. 1991; 173(18):5612-8. PMC: 208288. DOI: 10.1128/jb.173.18.5612-5618.1991. View

13.

Tomb J, El-Hajj H, Smith H . Nucleotide sequence of a cluster of genes involved in the transformation of Haemophilus influenzae Rd. Gene. 1991; 104(1):1-10. DOI: 10.1016/0378-1119(91)90457-m. View

14.

Barcak G, Chandler M, Redfield R, Tomb J . Genetic systems in Haemophilus influenzae. Methods Enzymol. 1991; 204:321-42. DOI: 10.1016/0076-6879(91)04016-h. View

15.

BUCHANAN C, Ling M . Isolation and sequence analysis of dacB, which encodes a sporulation-specific penicillin-binding protein in Bacillus subtilis. J Bacteriol. 1992; 174(6):1717-25. PMC: 205771. DOI: 10.1128/jb.174.6.1717-1725.1992. View

16.

Dorocicz I, Williams P, Redfield R . The Haemophilus influenzae adenylate cyclase gene: cloning, sequence, and essential role in competence. J Bacteriol. 1993; 175(22):7142-9. PMC: 206855. DOI: 10.1128/jb.175.22.7142-7149.1993. View

17.

Zulty J, Barcak G . Structural organization, nucleotide sequence, and regulation of the Haemophilus influenzae rec-1+ gene. J Bacteriol. 1993; 175(22):7269-81. PMC: 206870. DOI: 10.1128/jb.175.22.7269-7281.1993. View

18.

Clifton S, McCarthy D, Roe B . Sequence of the rec-2 locus of Haemophilus influenzae: homologies to comE-ORF3 of Bacillus subtilis and msbA of Escherichia coli. Gene. 1994; 146(1):95-100. DOI: 10.1016/0378-1119(94)90840-0. View

19.

Wren B, Henderson J, Ketley J . A PCR-based strategy for the rapid construction of defined bacterial deletion mutants. Biotechniques. 1994; 16(6):994-6. View

20.

Williams P, Bannister L, Redfield R . The Haemophilus influenzae sxy-1 mutation is in a newly identified gene essential for competence. J Bacteriol. 1994; 176(22):6789-94. PMC: 197045. DOI: 10.1128/jb.176.22.6789-6794.1994. View