lcrH, a gene necessary for virulence of Yersinia pestis and for the normal response of Y. pestis to ATP and calcium.
We are investigating the functions of the three proteins encoded by the V operon (lcrGVH) of the low-calcium response virulence plasmid pCD1 of Yersinia pestis KIM5. The purpose of this study was to define the role of the 18-kilodalton protein encoded by lcrH, the third gene of the V operon. Using marker exchange mutagenesis, we constructed a Y. pestis mutant that failed to express the LcrH protein. This LcrH- mutant was "ATP blind" in that it failed to show altered growth and V-antigen expression at 37 degrees C when 18 mM ATP was present. It also showed only a partial response to 2.5 mM Ca2+. The parental Y. pestis strain showed full growth yield at 37 degrees C and depressed expression of V antigen and of yop (yersinial pCD1-encoded outer membrane protein) genes in response to ATP or Ca2+. In contrast, the LcrH- mutant failed to grow at 37 degrees C in the presence of ATP and showed only limited growth when Ca2+ was present. V-antigen expression in the mutant was not depressed by ATP and only partially depressed by Ca2+. These findings show that LcrH is necessary for the normal response of Y. pestis to ATP and that LcrH contributes to Ca2+ responsiveness. The mutant also showed abnormal yopJ expression, indicating that LcrH also is necessary for normal yop regulation. The LcrH- mutant was avirulent in mice, probably because of its compromised growth at 37 degrees C. These findings indicate that the responses of Y. pestis to ATP and Ca2+ are distinct and that lcrH encodes a protein that is an important mediator of Ca2+ and ATP regulation of pCD1-encoded virulence determinant(s) in Y. pestis.
Price
S B
SB
Department of Microbiology and Immunology, Chandler Medical Center, University of Kentucky, Lexington 40536.
Straley
S C
SC
D000255:Adenosine Triphosphate / Q000494:pharmacology; D000942:Antigens, Bacterial / Q000235:genetics*; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics*; D002118:Calcium / Q000494:pharmacology; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D004252:DNA Mutational Analysis; D015180:Electrophoresis, Gel, Two-Dimensional; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005809:Genes, Regulator; D007526:Isoelectric Point; D008970:Molecular Weight; D009876:Operon; D015183:Restriction Mapping; D015010:Yersinia pestis / Q000235:genetics* / Q000254:growth & development / Q000472:pathogenicity
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D000255:Adenosine Triphosphate; D015500:Chloramphenicol O-Acetyltransferase; D002118:Calcium