Sequence analysis of the chromosomal region around and within the V-1-encoding gene of Mycoplasma pulmonis: Evidence for DNA inversion as a mechanism for V-1 variation

被引:41
作者
Simmons, WL
Cao, ZH
Glass, JI
Simecka, JW
Cassell, GH
Watson, HL
机构
[1] UNIV ALABAMA, SCH MED, DEPT MICROBIOL, BBRB 618, BIRMINGHAM, AL 35294 USA
[2] UNIV ALABAMA, SCH DENT, DEPT MICROBIOL, BIRMINGHAM, AL 35294 USA
关键词
D O I
10.1128/IAI.64.2.472-479.1996
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
Although the variation of V-1 antigens of Mycoplasma pulmonis has been correlated with variable expression of the cytadherence properties of this organism and has been implicated as a virulence determining factor in M. pulmonis-induced murine respiratory disease, the precise function of these antigens remains unknown. We have cloned and characterized genes encoding V-1 from two M. pulmonis UAB CT V-1 variants that differ in hemadsorption properties. A comparison of the nucleotide sequences revealed that these two variant genes were identical in the 5'-most 724 nucleotides. Regions of extensive divergence that contained repeated sequences were found 3' to this conserved region. On the basis of their deduced amino acid sequences, one variant expressed a V-1 protein of 91.2 kDa presumptively containing 40 repeats of 17 amino acids and the other expressed a protein of 27.4 kDa containing 2 direct, noncontiguous 9-amino-acid repeats. These general properties, as well as the presence of a prokaryotic lipoprotein acylation sequence (L-X-Y-C), indicated that the genes encoding V-1 were similar in structure to genes encoding other mycoplasma surface lipoproteins. Further analysis of sequences flanking these genes revealed that these variants arose via an inversion event which provided an interchange of the two variable regions with a single conserved region. Northern (RNA) analyses using oligonucleotide probes specific for the two different variable regions as well as for the conserved region of these genes and immunoblot analyses using rabbit polyclonal antibodies specific for synthetic peptides derived from the sequences of the different variable regions indicated that DNA inversion acted as a switch which allowed only one of the two different genes to be expressed at any given time. This inversion model clearly provides a mechanism by which M. pulmonis can alter its surface architecture and also strongly suggests that the as-yet-undefined function of V-1 resides in the variable carboxy region of these proteins.
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页码:472 / 479
页数:8
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