Sequence analysis of a 7.494 kb DNA fragment from megaplasmid 2 of
Rhizobium meliloti 2011 involved in exopolysaccharide I (EPS 1) biosynthesis revealed the presence of five
exo genes designated
exoA,
exoM,
exoN,
exoO, and
exoP. ExoN was found to show strong homology to a UDP-glucose pyrophosphorylase from
Acetobacter xylinum, whereas ExoO displayed weak homologies to the NodC proteins from
R. meliloti and
R. loti. Supprisingly, different mutations in
exoP resulted in divergent phenotypes. One
exoP mutant was able to establish an effective symbiosis with alfalfa, although no EPS I polymer could be detected. In contrast, other
exoP mutations prevented the formation of an effective symbiosis. The transcriptional organization of the
exoA-
exoP gene region has been analysed in conjunction with the
exoH,
exoK and
exoL genes. Using
exo-
lacZ transcription fusions in association with plasmid integration mutagenesis a strong promoter was identified upstream of
exoH, which is able to direct transcription of the whole
exoHKLAMONP gene cluster. A much weaker promoter upstream of
exoL was found to be involved in the transcription of the
exoLAMONP genes. In addition, weak promoters were identified upstream of
exoK,
exoA,
exoN and
exoP.
Key words
Rhizobium meliloti
- Exopolysaccharide biosynthesis - UDP-glucose pyrophosphorylase - Glucosyltransferase - Promoter mapping
Communicated by A. Kondorosi