Transcriptional regulation of the ospAB and ospC promoters from Borrelia burgdorferi.
OspA, OspB and OspC are the major outer surface proteins of Borrelia burgdorferi that are differentially synthesized in response to environmental conditions, including culture temperature. We found that DNA was more negatively supercoiled in B. burgdorferi cultures grown at 23 degrees C compared with cultures grown at 35-37 degrees C. We examined the regulation of ospAB and ospC transcription by temperature and DNA supercoiling. DNA supercoiling was relaxed by adding coumermycin A1, an antibiotic that inhibits DNA gyrase. Syntheses of the major outer surface proteins, expression of the ospA and ospC genes and the activities of the ospAB operon and ospC gene promoters were assayed. ospA product levels decreased, whereas ospC product levels increased after shifting from 23 degrees C to 35 degrees C or after adding coumermycin A1. In addition, OspC synthesis was higher in a gyrB mutant than in wild-type B. burgdorferi. Promoter activity was quantified using cat reporter fusions. Increasing temperature or relaxing supercoiled DNA resulted in a decrease in ospAB promoter activity in B. burgdorferi, but not in Escherichia coli, as well as an increase in ospC promoter activity in both bacteria. ospC promoter activity was increased in an E. coli gyrB mutant with an attenuated DNA supercoiling phenotype. These results suggest that B. burgdorferi senses environmental changes in temperature by altering the level of DNA supercoiling, which then affects the expression of the ospAB operon and the ospC gene. This implies that DNA supercoiling acts as a signal transducer for environmental regulation of outer surface protein synthesis.
Alverson
Janet
J
Division of Biological Sciences, The University of Montana, 32 Campus Dr. # 4824, Missoula 59812-4824, USA.
Bundle
Sharyl F
SF
Sohaskey
Charles D
CD
Lybecker
Meghan C
MC
Samuels
D Scott
DS
D049933:Aminocoumarins; D000942:Antigens, Bacterial / Q000378:metabolism; D000954:Antigens, Surface / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D001428:Bacterial Vaccines; D025065:Borrelia burgdorferi / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D003374:Coumarins / Q000494:pharmacology; D003470:Culture Media; D004278:DNA, Superhelical / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial; D008074:Lipoproteins; D011401:Promoter Regions, Genetic / Q000235:genetics*; D015398:Signal Transduction; D013696:Temperature; D014158:Transcription, Genetic
D049933:Aminocoumarins; D000942:Antigens, Bacterial; D000954:Antigens, Surface; D001425:Bacterial Outer Membrane Proteins; D001428:Bacterial Vaccines; D003374:Coumarins; D003470:Culture Media; D004278:DNA, Superhelical; D008074:Lipoproteins; C072045:OspA protein; C079616:OspC protein; C077030:OspB protein, Borrelia burgdorferi; C004628:coumermycin