Marrs, Carl F. and Rozsa, Frank W. and Hackel, Meredith and Stevens, Scott P. and Glasgow, Anna C. (1990) Identification, cloning, and sequencing of piv, a new gene involved in inverting the pilin genes of Moraxella lacunata. Journal of Bacteriology, 172 (8). pp. 4370-4377. ISSN 0021-9193 http://resolver.caltech.edu/CaltechAUTHORS:20120509-105143727
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Moraxella lacunata is a bacterium that is a causative agent of human conjunctivitis and keratitis. We have previously cloned the Q and I pilin (formerly called β and α pilin) genes of Moraxella bovis and determined that an inversion of 2 kilobases (kb) of DNA determines which pilin gene is expressed. Using an M. bovis pilin gene as a hybridization probe to screen a λ ZAP library of M. lacunata DNA, we have isolated a clone that not only contains the entire type 4 pilin gene inversion region of M. lacunata but inverts the 2-kb region on a plasmid subclone (pMxL1) in Escherichia coli. Deletion derivatives of pMxL1 yielded some plasmids that still had the entire inversion region but were phase locked into one or the other of the two potential orientations. Similarly, insertions of a 2-kb streptomycin-resistant element (Ω) within some regions outside of the inversion also resulted in phase-locked plasmids. These deletions and insertions thus localize a probable invertase necessary for the inversion event. The region was sequenced, and an open reading frame with over 98% DNA sequence homology to an open reading frame that we previously found in M. bovis and called ORF2 appeared to be a strong candidate for the invertase. This conclusion was confirmed when a plasmid containing the M. bovis ORF2 supplied, in trans, the inversion function missing from one of the M. lacunata phase-locked inversion mutants. We have named these putative invertase genes piv_(ml) (pilin inversion of M. lacunata) and piv_(mb) (pilin inversion of M. bovis). Despite previously noted sequence similarities between the M. bovis sites of inversion and those of the Hin family of invertible segments and a 60-base-pair region within the inversion with 50% sequence similarity to the cin recombinational enhancer, there is no significant sequence similarity of the Piv invertases to the Hin family of invertases.
|Additional Information:||© 1990 American Society for Microbiology. Received 20 February 1990; Accepted 23 May 1990. This work was supported by Public Health Service grant BM-1 1R01EY07125 from the National Eye Institute. A.C.G. was supported by the California Division of the American Cancer Society.|
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|Deposited By:||Tony Diaz|
|Deposited On:||09 May 2012 22:20|
|Last Modified:||26 Dec 2012 15:10|
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