Transcription of Proteus mirabilis flaAB.
Proteus mirabilis, a Gram-negative urinary tract pathogen, has two highly homologous, tandemly arranged flagellin-encoding genes, flaA and flaB. flaA is transcribed from a sigma(28) promoter, while flaB is a silent allele. Previous studies have demonstrated the presence of a family of hybrid flagellin genes, referred to as flaAB. These genes are composed of the 5' end of flaA and the 3' end of flaB, and are produced through excision of the intervening DNA between the two genes. Although the existence of flaAB DNA has been documented, it was not known if transcription of flaAB occurs in wild-type P. mirabilis. In this study, proof of flaAB transcription was obtained from a combination of RNA dot-blots and RT-PCR assays using specific primers and probes for flaAB and flaA. The RNA data were further supported by the demonstration of phenotypic switching of the locus using a FlaAB-detector strain. The results show that flaAB mRNA is transcribed and is 1/64 as abundant as flaA in the population of wild-type cells, suggesting that flaAB constitutes 1.0-1.5 % of the total flagellin message. Nucleotide sequence analysis of flaAB products produced by RT-PCR from the wild-type confirms previous reports of a variable fusion site between flaA and flaB resulting in a hybrid flagellin transcript. These data support the hypothesis that the production of FlaAB is integral to the physiology of P. mirabilis.
Manos
Jim
J
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 East Pratt Street, Baltimore, MD 21202, USA.
Belas
Robert
R
Center of Marine Biotechnology, University of Maryland Biotechnology Institute, 701 East Pratt Street, Baltimore, MD 21202, USA.
D000595:Amino Acid Sequence; D019677:Artificial Gene Fusion; D001426:Bacterial Proteins / Q000235:genetics; D001483:Base Sequence; D005408:Flagellin / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D007763:Lac Operon; D008969:Molecular Sequence Data; D009693:Nucleic Acid Hybridization; D011513:Proteus mirabilis / Q000235:genetics*; D012329:RNA, Bacterial / Q000032:analysis / Q000302:isolation & purification; D012333:RNA, Messenger / Q000032:analysis / Q000302:isolation & purification; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics / Q000378:metabolism
D001426:Bacterial Proteins; D012329:RNA, Bacterial; D012333:RNA, Messenger; D005408:Flagellin; C067138:flaA protein, bacteria; C416532:flaB flagellin; D001616:beta-Galactosidase