Verification and dissection of the ospC operator by using flaB promoter as a reporter in Borrelia burgdorferi.
The Lyme disease spirochete Borrelia burgdorferi must repress expression of outer surface protein C (OspC) to effectively evade specific humoral immunity and to establish persistent infection. This ability largely relies upon a regulatory element, the only operator that has been reported in spirochetal bacteria. Immediately upstream of the ospC promoter, two sets of inverted repeats (IRs) constitute small and large palindromes, in which the right IR of the large palindrome contains the left IR of the small one, and may collectively function as the ospC operator. In the study, the large palindrome with or without the small IR was fused with an flaB promoter, which was used to drive expression of a promoterless ospC copy as a reporter gene, and introduced into OspC-deficient B. burgdorferi. The presence of the large palindrome alone significantly reduced ospC expression driven by the fused flaB promoter in the joint tissue of severe combined immunodeficiency (SCID) mice, and rescued spirochetes from elimination by passively transferred OspC antibody in infected SCID mice and specific immune responses elicited in immunocompetent mice, confirming a function of the IRs as an operator. Inclusion of the small IR further enhanced the ability of the large palindrome to reduce the activity of the fused flaB promoter, indicating that the small IR is a part of the operator. Taken together, the study led to successful verification and dissection of the ospC operator.
Xu
Qilong
Q
Department of Pathobiological Sciences, Louisiana State University, Baton Rouge, LA 70803, USA.
McShan
Kristy
K
Liang
Fang Ting
FT
D000818:Animals; D000942:Antigens, Bacterial / Q000235:genetics* / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D025065:Borrelia burgdorferi / Q000235:genetics* / Q000276:immunology / Q000378:metabolism; D005408:Flagellin / Q000235:genetics*; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D006801:Humans; D008193:Lyme Disease / Q000276:immunology / Q000382:microbiology*; D051379:Mice; D016513:Mice, SCID; D008969:Molecular Sequence Data; D009875:Operator Regions, Genetic / Q000276:immunology; D011401:Promoter Regions, Genetic; D012091:Repetitive Sequences, Nucleic Acid
D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; C079616:OspC protein; D005408:Flagellin; C416532:flaB flagellin