Localization of plasmid loci necessary for the entry of Shigella flexneri into HeLa cells, and characterization of one locus encoding four immunogenic polypeptides.
We have previously cloned a 44 kb fragment from the virulence plasmid of Shigella flexneri serotype 5 strain M90T which is capable of restoring invasiveness to an avirulent, plasmidless mutant. This report presents a genetic and physical analysis of Tn5 mutations in recombinant clone pHS4108. Tn5 mutagenesis allowed identification of at least five regions implicated in the entry phenotype. These regions were located on a 20 kb portion of pHS4108. Expression of the insertion mutants was studied by immunoblots using the serum of a convalescent monkey infected by S. flexneri 2a, which recognized four plasmid-associated polypeptides. We propose that the four immunogenic polypeptides, a, b, c, and d, are encoded an operon.
Baudry
B
B
Service des Entérobactéries, Unité INSERM 199, Institut Pasteur, Paris, France.
Maurelli
A T
AT
Clerc
P
P
Sadoff
J C
JC
Sansonetti
P J
PJ
D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004251:DNA Transposable Elements; D005798:Genes, Bacterial; D006367:HeLa Cells; D006801:Humans; D009154:Mutation; D010455:Peptides / Q000235:genetics*; D010957:Plasmids; D014176:Protein Biosynthesis; D012763:Shigella flexneri / Q000235:genetics* / Q000472:pathogenicity
D004251:DNA Transposable Elements; D010455:Peptides