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The low-Ca2+ -response (LCR) plasmid pCD1 of the plague agentYersinia pestis KIM5 was sequenced and analyzed for its genetic structure. pCD1 (70,509 bp) has an IncFIIA-like replicon and a SopABC-like partition region. We have assigned 60 apparently intact open reading frames (ORFs) that are not contained within transposable elements. Of these, 47 are proven or possible members of the LCR, a major virulence property of human-pathogenicYersinia spp., that had been identified previously in one or more ofY. pestis or the enteropathogenic yersiniaeYersinia enterocolitica andYersinia pseudotuberculosis . Of these 47 LCR-related ORFs, 35 constitute a continuous LCR cluster. The other LCR-related ORFs are interspersed among three intact insertion sequence (IS) elements (IS100 and two new IS elements, IS1616 and IS1617 ) and numerous defective or partial transposable elements. Regional variations in percent GC content and among ORFs encoding effector proteins of the LCR are additional evidence of a complex history for this plasmid. Our analysis suggested the possible addition of a new Syc- and Yop-encoding operon to the LCR-related pCD1 genes and gave no support for the existence of YopL. YadA likely is not expressed, as was the case forY. pestis EV76, and the gene for the lipoprotein YlpA found inY. enterocolitica likely is a pseudogene inY. pestis . TheyopM gene is longer than previously thought (by a sequence encoding two leucine-rich repeats), the ORF upstream ofypkA-yopJ is discussed as a potential Syc gene, and a previously undescribed ORF downstream ofyopE was identified as being potentially significant. Eight other ORFs not associated with IS elements were identified and deserve future investigation into their functions.

DNA Sequencing and Analysis of the Low-Ca2+-Response Plasmid pCD1 ofYersinia pestisKIM5

DNA Sequencing and Analysis of the Low-Ca²⁺-Response Plasmid pCD1 ofYersinia pestisKIM5