Molecular characterization of a 40 kDa OmpC-like porin from Serratia marcescens.
An oligonucleotide that encodes the N-terminal portion of a 41 kDa porin of Serratia marcescens was used to probe S. marcescens UOC-51 genomic DNA. An 11 kb EcoRI fragment which hybridized with the oligonucleotide was subcloned into Escherichia coli, examined for expression, and sequenced. The product expressed by the cloned gene was 40 kDa. The nucleotide sequence has an ORF of 1.13 kb. When the deduced amino acid sequence was aligned and compared to other enterobacterial porins the cloned S. marcescens porin most closely resembled E. coli OmpC. Although we did not detect osmoregulation or thermoregulation of any porins in S. marcescens UOC-51, sequences analogous to the E. coli osmoregulator OmpR-binding regions are seen upstream to the cloned gene. We examined the regulation of the S. marcescens porin in E. coli and found that its expression increased in a high salt environment. A micF gene, whose transcriptional product functions to inhibit synthesis of OmpF by hybridizing with the ompF transcript, was also seen upstream of the S. marcescens ompC. An alignment with the E. coli micF gene revealed that the functional region of the S. marcescens micF gene is conserved. Based on the results obtained we have determined that S. marcescens UOC-51 produces a 40 kDa porin similar to the E. coli OmpC porin.
Hutsul
J A
JA
Department of Microbiology, University of Manitoba, Winnipeg, Canada.
Worobec
E
E
D000595:Amino Acid Sequence; D001425:Bacterial Outer Membrane Proteins / Q000096:biosynthesis / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D016384:Consensus Sequence; D004926:Escherichia coli / Q000187:drug effects; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D018272:Porins / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D012462:Saline Solution, Hypertonic / Q000494:pharmacology; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D012706:Serratia marcescens / Q000737:chemistry* / Q000235:genetics; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D015345:Oligonucleotide Probes; D018272:Porins; D011993:Recombinant Fusion Proteins; D012462:Saline Solution, Hypertonic