The gene encoding a Prevotella loescheii lectin-like adhesin contains an interrupted sequence which causes a frameshift.
We cloned and sequenced the Prevotella loescheii gene plaA, which encodes a lectin-like adhesin that mediates the coaggregation of P. loescheii 1295 with Streptococcus oralis 34. A probe derived from the N-terminal amino acid sequence of the purified adhesin was used to identify the plaA gene from a P. loescheii genomic library constructed in lambda GEM-11. Sequence analysis of plaA indicates that the initial translation product contains a 22-amino-acid leader. The reading frame of the plaA gene is interrupted after amino acid 28 of the mature protein by a TAA termination codon. Amplification of the P. loescheii genomic DNA in the region surrounding this codon by the polymerase chain reaction followed by DNA sequencing of the cloned DNA fragment established that this stop codon was not an experimental artifact. A frameshift beginning 29 bp downstream of the ochre terminator was required to access the only large open reading frame in the gene. Amino acid sequences of six purified peptides derived by limited proteolysis of adhesin with endoproteinase Lys-C matched the downstream amino acid sequence derived by translation of the large open reading frame. The gene coding sequence of 2.4 kb contains sufficient information for the synthesis of an 89-kDa protein. A putative rho-independent terminator (delta G = -25.5 kcal/mol [ca. -107 kJ/mol]) was detected 38 bp downstream from the plaA stop codon.
Manch-Citron
J N
JN
Laboratory of Microbial Ecology, National Institute of Dental Research, Food and Drug Administration, Bethesda, Maryland 20892.
Allen
J
J
Moos
M
M
London
J
J
D018829:Adhesins, Bacterial; D000595:Amino Acid Sequence; D001422:Bacterial Adhesion / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D001439:Bacteroides / Q000235:genetics*; D001483:Base Sequence; D015139:Blotting, Southern; D002151:Calorimetry; D003001:Cloning, Molecular; D003062:Codon; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics; D016368:Frameshift Mutation; D005798:Genes, Bacterial; D015698:Genomic Library; D037102:Lectins / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D015345:Oligonucleotide Probes; D016366:Open Reading Frames; D010957:Plasmids; D011487:Protein Conformation; D015183:Restriction Mapping; D017386:Sequence Homology, Amino Acid; D013728:Terminator Regions, Genetic
D018829:Adhesins, Bacterial; D001426:Bacterial Proteins; D003062:Codon; D004269:DNA, Bacterial; D037102:Lectins; D015345:Oligonucleotide Probes; C063910:adhesin, Bacteroides; C086517:plaA protein, Prevotella loescheii