In vitro Tn7 mutagenesis of Haemophilus influenzae Rd and characterization of the role of atpA in transformation

M. L. Gwinn, A. E. Stellwagen, Nancy L Craig, J. F. Tomb, H. O. Smith

Research output: Contribution to journalArticle

Abstract

Haemophilus influenzae Rd is a gram-negative bacterium capable of natural DNA transformation. The competent state occurs naturally in late exponential growth or can be induced by a nutritional downshift or by transient anaerobiosis. The genes cya, crp, topA, and sxy (tfoX) are known to function in the regulation of competence development. The phosphoenolpyruvate:carbohydrate phosphotransferase system functions to maintain levels of cyclic AMP necessary for competence development but is not directly involved in regulation. The exact signal(s) for competence and the genes that mediate the signal(s) are still unknown. In an effort to find additional regulatory genes, H. influenzae Rd was mutated by using an in vitro Tn7 system and screened for mutants with a reduced ability to induce the competence-regulatory gene. comA. Insertions in atpA, a gene coding for the α subunit of the F1 cytoplasmic domain of the ATP synthase, reduce transformation frequencies about 20-fold and cause a significant reduction in expression of competence-regulatory genes, while the expression of constitutive competence genes is only minimally affected. In addition, we found that an insertion in atpB, which encodes the a subunit of the F0 membrane-spanning domain, has a similar effect on transformation frequencies.

Original languageEnglish (US)
Pages (from-to)7315-7320
Number of pages6
JournalJournal of Bacteriology
Volume179
Issue number23
StatePublished - Dec 1997

Fingerprint

Haemophilus influenzae
Mutagenesis
Mental Competency
Regulator Genes
Genes
Anaerobiosis
Phosphoenolpyruvate
Aptitude
Coma
Gram-Negative Bacteria
Cyclic AMP
In Vitro Techniques
Phosphotransferases
Adenosine Triphosphate
Carbohydrates
Gene Expression
Membranes
DNA
Growth

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

In vitro Tn7 mutagenesis of Haemophilus influenzae Rd and characterization of the role of atpA in transformation. / Gwinn, M. L.; Stellwagen, A. E.; Craig, Nancy L; Tomb, J. F.; Smith, H. O.

In: Journal of Bacteriology, Vol. 179, No. 23, 12.1997, p. 7315-7320.

Research output: Contribution to journalArticle

Gwinn, M. L. ; Stellwagen, A. E. ; Craig, Nancy L ; Tomb, J. F. ; Smith, H. O. / In vitro Tn7 mutagenesis of Haemophilus influenzae Rd and characterization of the role of atpA in transformation. In: Journal of Bacteriology. 1997 ; Vol. 179, No. 23. pp. 7315-7320.
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