Membrane topology analysis of the Bacillus subtilis BofA protein involved in pro-sigma K processing.
The Bacillus subtilis BofA protein is involved in regulation of pro-sigma K processing in the mother cell during the late stages of sporulation. A computer analysis of the BofA amino acid sequence indicates that it is an integral membrane protein. To determine the membrane topology of the protein, a series of gene fusions of bofA with lacZ or phoA reporter genes in Escherichia coli were analysed. A BofA topological model with two membrane-spanning segments, and with the N- and the C-terminal domains located in the region between the inner and outer membranes surrounding the forespore is presented. The analysis of different modifications of the last five amino acid residues of the BofA protein, obtained by PCR site-directed mutagenesis, suggests a possible role of the C-terminal domain in the regulation of pro-sigma K processing.
Varcamonti
Mario
M
Istituto di Scienze delĺAlimentazione, Consiglio Nazionale delle Ricerche, via Roma, 83100 Avellino, Italy.
Marasco
Rosangela
R
Dipartimento di Chimica, Università degli Studi di Salerno, via Ponte Don Melillo, 84084 Fisciano (Sa), Italy.
Maurilio
De Felice
F
Sezione di Microbiologia, Dipartimento di Fisioiogia Generale ed Ambientale, Universitá Federico II, via Mezzocannone 16, 80134 Naples, Italy.
Sacco
Margherita
M
Istituto Internazionale di Genetica e Biofisica, Consiglio Nazionale delle Ricerche, via G. Marconi 10, 80125 Naples, Italy.
D000469:Alkaline Phosphatase / Q000235:genetics; D000595:Amino Acid Sequence; D001412:Bacillus subtilis / Q000737:chemistry* / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D002451:Cell Compartmentation; D007763:Lac Operon; D008565:Membrane Proteins / Q000737:chemistry* / Q000235:genetics; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011498:Protein Precursors / Q000378:metabolism; D011499:Protein Processing, Post-Translational; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D013171:Spores, Bacterial / Q000737:chemistry; D014157:Transcription Factors / Q000378:metabolism
D001426:Bacterial Proteins; D008565:Membrane Proteins; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D014157:Transcription Factors; C084190:sigma K; C074423:bofA protein, Bacillus subtilis; D000469:Alkaline Phosphatase