The dspA gene product of the cyanobacterium Synechocystis sp. strain PCC 6803 influences sensitivity to chemically different growth inhibitors and has amino acid similarity to histidine protein kinases.
We have cloned and sequenced a gene of the cyanobacterium Synechocystis sp. strain PCC 6803 named dspA (encoding drug sensory protein A; DspA), mutations in which result in cross-resistance to the herbicides difunon and diuron, as well as to the calmodulin antagonists chlorpromazine and trifluoperazine. The dspA gene encodes a polypeptide of 663 amino acids with a predicted molecular mass of 74.5 kDa. The molecular nature of two mutations in the dspA gene leading to the cross-resistance has been determined. Targeted mutagenesis of the dspA gene was performed using a kanamycin-resistance gene cartridge. Resulting mutant strains were checked for resistance to difunon and chlorpromazine and showed cross-resistance to both agents. The C-terminal portion of the deduced amino acid sequence of DspA shares significant similarity with the conserved region of histidine protein kinases (HPKs). Hydrophobicity analysis of the amino acid sequence of DspA indicated the existence of two hydrophobic regions in the N-terminal portion that are characteristic of the bacterial sensory HPK family. We suggest that protein DspA is a HPK involved in chemical sensing.
Bartsevich
V V
VV
N. Vavilov Institute of General Genetics, Moscow, Russia.
Shestakov
S V
SV
D000595:Amino Acid Sequence; D001483:Base Sequence; D002746:Chlorpromazine / Q000494:pharmacology; D003001:Cloning, Molecular; D000458:Cyanobacteria / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics; D004237:Diuron / Q000494:pharmacology; D004352:Drug Resistance, Microbial / Q000235:genetics; D005798:Genes, Bacterial; D006540:Herbicides / Q000494:pharmacology; D000071677:Histidine Kinase; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011494:Protein Kinases / Q000235:genetics*; D017386:Sequence Homology, Amino Acid; D014268:Trifluoperazine / Q000494:pharmacology
D004269:DNA, Bacterial; D006540:Herbicides; D014268:Trifluoperazine; D004237:Diuron; D011494:Protein Kinases; D000071677:Histidine Kinase; D002746:Chlorpromazine