A novel CRP-dependent regulon controls expression of competence genes in Haemophilus influenzae.
Natural competence for DNA uptake is common among bacteria but its evolutionary function is controversial. Resolving the dispute requires a detailed understanding of both how cells decide to take up DNA and how the DNA is processed during and after uptake. We have used whole-genome microarrays to follow changes in gene expression during competence development in wild-type Haemophilus influenzae cells, and to characterize dependence of competence-induced transcription on known regulatory factors. This analysis confirmed the existence of a postulated competence regulon, characterized by a promoter-associated 22 bp competence regulatory element (CRE) closely related to the cAMP receptor protein (CRP) binding consensus. This CRE regulon contains 25 genes in 13 transcription units, only about half of which have been previously associated with competence. The new CRE genes encode a periplasmic ATP-dependent DNA ligase, homologs of SSB, RadC and the Bacillus subtilis DNA uptake protein ComEA, and eight genes of unknown function. Competence-induced transcription of genes in the CRE regulon is strongly dependent on cAMP, consistent with the known role of catabolite regulation in competence. Electrophoretic mobility-shift assays confirmed that CRE sequences are a new class of CRP-binding site. The essential competence gene sxy is induced early in competence development and is required for competence-induced transcription of CRE-regulon genes but not other CRP-regulated genes, suggesting that Sxy may act as an accessory factor directing CRP to CRE sites. Natural selection has united these 25 genes under a common regulatory mechanism. Elucidating this mechanism, and the functions of the genes, will provide a valuable window into the evolutionary function of natural competence.
Redfield
Rosemary J
RJ
Department of Zoology, University of British Columbia, Vancouver, Canada V6T 1Z4. redfield@interchange.ubc.ca
Cameron
Andrew D S
AD
Qian
Qing
Q
Hinds
J
J
Ali
Tahir R
TR
Kroll
J Simon
JS
Langford
Paul R
PR
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D003470:Culture Media / Q000494:pharmacology; D000242:Cyclic AMP / Q000494:pharmacology; D002373:Cyclic AMP Receptor Protein / Q000378:metabolism*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D024202:Electrophoretic Mobility Shift Assay; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects / Q000235:genetics*; D005798:Genes, Bacterial / Q000235:genetics*; D006193:Haemophilus influenzae / Q000187:drug effects / Q000235:genetics*; D020411:Oligonucleotide Array Sequence Analysis; D018085:Regulon / Q000235:genetics*; D020218:Response Elements / Q000235:genetics*; D015534:Trans-Activators / Q000235:genetics / Q000378:metabolism; D014169:Transformation, Bacterial / Q000235:genetics*
D001426:Bacterial Proteins; D003470:Culture Media; D002373:Cyclic AMP Receptor Protein; C093149:Tfox protein, Haemophilus influenzae; D015534:Trans-Activators; D000242:Cyclic AMP