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. 1992 Mar 1;89(5):1626–1630. doi: 10.1073/pnas.89.5.1626

The gene encoding cAMP receptor protein is required for competence development in Haemophilus influenzae Rd.

M S Chandler 1
PMCID: PMC48505  PMID: 1542653

Abstract

The Haemophilus influenzae Rd strain JG87 contains a single mini-Tn10kan insertion that causes a deficiency in the development of competence for genetic transformation. The DNA fragment containing this insertion mutation, as well as the wild-type locus, was cloned, mapped, and sequenced. The sequence contained an open reading frame for a protein of 224 amino acids with a predicted Mr of 25,152. The deduced protein sequence showed strong similarity to the Escherichia coli cAMP receptor protein. The E. coli crp gene cloned on a multicopy plasmid was shown to fully complement the competence-deficient phenotype of the mutant strain; thus, the H. influenzae gene was named crp. These results suggest that H. influenzae cAMP-cAMP receptor protein complex functions to regulate one or more promoters essential for the development of competence in H. influenzae Rd. Features of a gene upstream of H. influenzae crp that is homologous to the E. coli ttk gene are also described.

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Selected References

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  1. Aiba H. Autoregulation of the Escherichia coli crp gene: CRP is a transcriptional repressor for its own gene. Cell. 1983 Jan;32(1):141–149. doi: 10.1016/0092-8674(83)90504-4. [DOI] [PubMed] [Google Scholar]
  2. Aiba H., Fujimoto S., Ozaki N. Molecular cloning and nucleotide sequencing of the gene for E. coli cAMP receptor protein. Nucleic Acids Res. 1982 Feb 25;10(4):1345–1361. doi: 10.1093/nar/10.4.1345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Barcak G. J., Chandler M. S., Redfield R. J., Tomb J. F. Genetic systems in Haemophilus influenzae. Methods Enzymol. 1991;204:321–342. doi: 10.1016/0076-6879(91)04016-h. [DOI] [PubMed] [Google Scholar]
  4. Beattie K. L., Setlow J. K. Transformation-defective strains of Haemophilus influenzae. Nat New Biol. 1971 Jun 9;231(23):177–179. doi: 10.1038/newbio231177a0. [DOI] [PubMed] [Google Scholar]
  5. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Chandler M. S., Morrison D. A. Competence for genetic transformation in Streptococcus pneumoniae: molecular cloning of com, a competence control locus. J Bacteriol. 1987 May;169(5):2005–2011. doi: 10.1128/jb.169.5.2005-2011.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chandler M. S. New shuttle vectors for Haemophilus influenzae and Escherichia coli: P15A-derived plasmids replicate in H. influenzae Rd. Plasmid. 1991 May;25(3):221–224. doi: 10.1016/0147-619x(91)90016-p. [DOI] [PubMed] [Google Scholar]
  9. Cossart P., Gicquel-Sanzey B. Cloning and sequence of the crp gene of Escherichia coli K 12. Nucleic Acids Res. 1982 Feb 25;10(4):1363–1378. doi: 10.1093/nar/10.4.1363. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cossart P., Gicquel-Sanzey B. Regulation of expression of the crp gene of Escherichia coli K-12: in vivo study. J Bacteriol. 1985 Jan;161(1):454–457. doi: 10.1128/jb.161.1.454-457.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Cossart P., Groisman E. A., Serre M. C., Casadaban M. J., Gicquel-Sanzey B. crp genes of Shigella flexneri, Salmonella typhimurium, and Escherichia coli. J Bacteriol. 1986 Aug;167(2):639–646. doi: 10.1128/jb.167.2.639-646.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Danner D. B., Deich R. A., Sisco K. L., Smith H. O. An eleven-base-pair sequence determines the specificity of DNA uptake in Haemophilus transformation. Gene. 1980 Nov;11(3-4):311–318. doi: 10.1016/0378-1119(80)90071-2. [DOI] [PubMed] [Google Scholar]
  13. Ebright R. H., Cossart P., Gicquel-Sanzey B., Beckwith J. Mutations that alter the DNA sequence specificity of the catabolite gene activator protein of E. coli. Nature. 1984 Sep 20;311(5983):232–235. doi: 10.1038/311232a0. [DOI] [PubMed] [Google Scholar]
  14. GOODGAL S. H., HERRIOTT R. M. Studies on transformations of Hemophilus influenzae. I. Competence. J Gen Physiol. 1961 Jul;44:1201–1227. doi: 10.1085/jgp.44.6.1201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Goodgal S. H., Mitchell M. A. Sequence and uptake specificity of cloned sonicated fragments of Haemophilus influenzae DNA. J Bacteriol. 1990 Oct;172(10):5924–5928. doi: 10.1128/jb.172.10.5924-5928.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Herriott R. M., Meyer E. M., Vogt M. Defined nongrowth media for stage II development of competence in Haemophilus influenzae. J Bacteriol. 1970 Feb;101(2):517–524. doi: 10.1128/jb.101.2.517-524.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lundberg L. G., Karlström O. H., Nyman P. O. Isolation and characterization of the dut gene of Escherichia coli. II. Restriction enzyme mapping and analysis of polypeptide products. Gene. 1983 Apr;22(1):127–131. doi: 10.1016/0378-1119(83)90071-9. [DOI] [PubMed] [Google Scholar]
  18. Lundberg L. G., Thoresson H. O., Karlström O. H., Nyman P. O. Nucleotide sequence of the structural gene for dUTPase of Escherichia coli K-12. EMBO J. 1983;2(6):967–971. doi: 10.1002/j.1460-2075.1983.tb01529.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Marko M. A., Chipperfield R., Birnboim H. C. A procedure for the large-scale isolation of highly purified plasmid DNA using alkaline extraction and binding to glass powder. Anal Biochem. 1982 Apr;121(2):382–387. doi: 10.1016/0003-2697(82)90497-3. [DOI] [PubMed] [Google Scholar]
  20. Martinez E., Bartolomé 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 Aug 15;68(1):159–162. doi: 10.1016/0378-1119(88)90608-7. [DOI] [PubMed] [Google Scholar]
  21. McKay D. B., Steitz T. A. Structure of catabolite gene activator protein at 2.9 A resolution suggests binding to left-handed B-DNA. Nature. 1981 Apr 30;290(5809):744–749. doi: 10.1038/290744a0. [DOI] [PubMed] [Google Scholar]
  22. Morrison D. A. Transformation and preservation of competent bacterial cells by freezing. Methods Enzymol. 1979;68:326–331. doi: 10.1016/0076-6879(79)68023-0. [DOI] [PubMed] [Google Scholar]
  23. Okamoto K., Hara S., Bhasin R., Freundlich M. Evidence in vivo for autogenous control of the cyclic AMP receptor protein gene (crp) in Escherichia coli by divergent RNA. J Bacteriol. 1988 Nov;170(11):5076–5079. doi: 10.1128/jb.170.11.5076-5079.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Redfield R. J. sxy-1, a Haemophilus influenzae mutation causing greatly enhanced spontaneous competence. J Bacteriol. 1991 Sep;173(18):5612–5618. doi: 10.1128/jb.173.18.5612-5618.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schroeder C. J., Dobrogosz W. J. Cloning and DNA sequence analysis of the wild-type and mutant cyclic AMP receptor protein genes from Salmonella typhimurium. J Bacteriol. 1986 Aug;167(2):616–622. doi: 10.1128/jb.167.2.616-622.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Setlow J. K., Cabrera-Juárez E., Griffin K. Mechanism of acquisition of chromosomal markers by plasmids in Haemophilus influenzae. J Bacteriol. 1984 Nov;160(2):662–667. doi: 10.1128/jb.160.2.662-667.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Shine J., Dalgarno L. Determinant of cistron specificity in bacterial ribosomes. Nature. 1975 Mar 6;254(5495):34–38. doi: 10.1038/254034a0. [DOI] [PubMed] [Google Scholar]
  29. Stuy J. H., Walter R. B. Effect of glycerol on plasmid transfer in genetically competent Haemophilus influenzae. Mol Gen Genet. 1986 May;203(2):296–299. doi: 10.1007/BF00333969. [DOI] [PubMed] [Google Scholar]
  30. Tomb J. F., Barcak G. J., Chandler M. S., Redfield R. J., Smith H. O. Transposon mutagenesis, characterization, and cloning of transformation genes of Haemophilus influenzae Rd. J Bacteriol. 1989 Jul;171(7):3796–3802. doi: 10.1128/jb.171.7.3796-3802.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Weber I. T., Steitz T. A. Structure of a complex of catabolite gene activator protein and cyclic AMP refined at 2.5 A resolution. J Mol Biol. 1987 Nov 20;198(2):311–326. doi: 10.1016/0022-2836(87)90315-9. [DOI] [PubMed] [Google Scholar]
  32. Wilcox K. W., Smith H. O. Isolation and characterization of mutants of Haemophilus influenzae deficient in an adenosine 5'-triphosphate-dependent deoxyribonuclease activity. J Bacteriol. 1975 May;122(2):443–453. doi: 10.1128/jb.122.2.443-453.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Wise E. M., Jr, Alexander S. P., Powers M. Adenosine 3':5'-cyclic monophosphate as a regulator of bacterial transformation. Proc Natl Acad Sci U S A. 1973 Feb;70(2):471–474. doi: 10.1073/pnas.70.2.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zoon K. C., Habersat M., Scocca J. J. Multiple regulatory events in the development of competence for genetic transformation in Haemophilus influenzae. J Bacteriol. 1975 Dec;124(3):1607–1609. doi: 10.1128/jb.124.3.1607-1609.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zoon K. C., Habersat M., Scocca J. J. Synthesis of envelope polypeptides by Haemophilus influenzae during development of competence for genetic transformation. J Bacteriol. 1976 Jul;127(1):545–554. doi: 10.1128/jb.127.1.545-554.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zoon K. C., Scocca J. J. Constitution of the cell envelope of Haemophilus influenzae in relation to competence for genetic transformation. J Bacteriol. 1975 Aug;123(2):666–677. doi: 10.1128/jb.123.2.666-677.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. de Crecy-Lagard V., Glaser P., Lejeune P., Sismeiro O., Barber C. E., Daniels M. J., Danchin A. A Xanthomonas campestris pv. campestris protein similar to catabolite activation factor is involved in regulation of phytopathogenicity. J Bacteriol. 1990 Oct;172(10):5877–5883. doi: 10.1128/jb.172.10.5877-5883.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. de Crombrugghe B., Busby S., Buc H. Cyclic AMP receptor protein: role in transcription activation. Science. 1984 May 25;224(4651):831–838. doi: 10.1126/science.6372090. [DOI] [PubMed] [Google Scholar]
  39. el-Hajj H. H., Zhang H., Weiss B. Lethality of a dut (deoxyuridine triphosphatase) mutation in Escherichia coli. J Bacteriol. 1988 Mar;170(3):1069–1075. doi: 10.1128/jb.170.3.1069-1075.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]

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