1887

Abstract

A gene encoding a serine acetyltransferase (SAT) potentially involved in the biosynthesis of cysteine was identified ∼4 kb upstream of the previously described gene cluster that encodes an amino acid permease in strain 3841. The gene exhibits >40% identity to the family of SATs containing N-terminal extensions that have been described for other bacteria and plants. The ORF has three possible translation initiation sites which potentially encode polypeptides of 311, 277 and/or 259 amino acid residues, respectively. All three ORFs complemented the mutation in an cysteine auxotroph, strain JM39. Insertion of Tn into in the genome of (strain RU632) lowered SAT activity in crude extracts by >95%. However, RU632 was not a cysteine auxotroph, which suggests that possesses some redundancy in cysteine biosynthesis. Additional copies of could not be detected in the genome when the gene was used as a hybridization probe. Therefore it is possible that possesses an alternative pathway for cysteine biosynthesis which avoids -acetylserine. Strain RU632 was unaffected in its ability to nodulate , and the nodules were effective for N fixation (measured by CH reduction). Transcriptional activity of was determined by measuring the β-galactosidase arising from ::Tn fusions. Maximal levels of expression were observed during early exponential growth and were not influenced by the level of sulphur (supplied as sulphate). However, transcription was repressed by approximately twofold in ammonium-grown, as opposed to glutamate-grown, cultures. Repression by ammonium was not seen in a strain defective for .

Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-147-9-2553
2001-09-01
2020-03-28
Loading full text...

Full text loading...

/deliver/fulltext/micro/147/9/1472553a.html?itemId=/content/journal/micro/10.1099/00221287-147-9-2553&mimeType=html&fmt=ahah

References

  1. Altschul S. F., Madden T. L., Zhang J., Zhang Z., Miller W., Lipman D. J., Schäffer A. A.. 1997; Gapped blast and psi-blast: a new generation of protein database search programs. Nucleic Acids Res25:3389–3402[CrossRef]
    [Google Scholar]
  2. Beringer J. E.. 1974; R factor transfer in Rhizobium leguminosarum . J Gen Microbiol84:188–198[CrossRef]
    [Google Scholar]
  3. Bogdanova N., Hell R.. 1997; Cysteine synthesis in plants: protein-protein interactions of serine acetyltransferase from Arabidopsis thaliana . Plant J11:251–262[CrossRef]
    [Google Scholar]
  4. Cherest H., Surdin-Kerjan Y.. 1992; Genetic analysis of a new mutation conferring cysteine auxotrophy in Saccharomyces cerevisiae : updating of the sulfur metabolism pathway. Genetics130:51–58
    [Google Scholar]
  5. Corpet F.. 1988; Multiple sequence alignment with hierarchical clustering. Nucleic Acids Res16:10881–10890[CrossRef]
    [Google Scholar]
  6. Denk D., Böck A.. 1987; l-Cysteine biosynthesis in Escherichia coli : nucleotide sequence and expression of the serine acetyl transferase ( cysE ) gene from the wild-type and a cysteine excreting mutant. J Gen Microbiol133:515–525
    [Google Scholar]
  7. Evans D. J., Jones R., Woodley P. R., Wilborn J. R., Robson R. L.. 1991; Nucleotide sequence and genetic analysis of the Azotobacter chroococcum nifUSVWZM cluster, including a new gene ( nifP ) which encodes a serine acetyl transferase. J Bacteriol173:5457–5469
    [Google Scholar]
  8. Foglino M., Borne F., Bally M., Ball G., Patte J. C.. 1995; A direct thiolation pathway is used for methionine biosynthesis in Pseudomonas aeruginosa . Microbiology141:431–439[CrossRef]
    [Google Scholar]
  9. Glenn A. R., Poole P. S., Hudman J. F.. 1980; Succinate uptake by free-living and bacteroid forms of Rhizobium leguminosarum . J Gen Microbiol119:267–271
    [Google Scholar]
  10. Hanahan D.. 1983; Studies on transformation of Escherichia coli with plasmids. J Mol Biol166:557–580[CrossRef]
    [Google Scholar]
  11. Howarth J. R., Roberts M. A., Wray J. L.. 1997; Cysteine biosynthesis in higher plants: a new member of the Arabidopsis thaliana serine acetyltransferase small gene-family obtained by functional complementation of an Escherichia coli cysteine auxotroph. Biochim Biophys Acta1350:123–127[CrossRef]
    [Google Scholar]
  12. Huang T. C., Lin R. F., Chu M. K., Chen H. M.. 1999; Organization and expression of nitrogen-fixation genes in the aerobic nitrogen-fixing unicellular cyanobacterium Synechococcus sp. strain RF-1. Microbiology145:743–753[CrossRef]
    [Google Scholar]
  13. Jones-Mortimer M. C.. 1968; Positive control of sulphate reduction in Escherichia coli . The nature of the pleiotropic cysteineless mutants of E. coli K12. Biochem J110:597–602
    [Google Scholar]
  14. Kredich N. M.. 1987; Biosynthesis of cysteine. In Escherichia coli and Salmonella typhimurium: Cellular and Molecular Biology pp419–428 Edited by Neidhardt F. C.. and others Washington, DC: American Society for Microbiology;
    [Google Scholar]
  15. Kredich N. M., Tomkins G. M.. 1966; The enzymic synthesis of l-cysteine in Escherichia coli and Salmonella typhimurium. J Biol Chem241:4955–4965
    [Google Scholar]
  16. Mead D. A., Long S., Ruvkin G., Brown S., Ausubel F.. 1982; Physical and genetic characterization of symbiotic and auxotrophic mutants of Rhizobium meliloti . J Bacteriol149:114–122
    [Google Scholar]
  17. Mead D. A., Szczesna-Skorupa E., Kemper B.. 1986; Single-stranded DNA ‘blue’ promoter plasmids: a versatile tandem promoter system for cloning and protein engineering. Protein Eng1:67–74
    [Google Scholar]
  18. Miller J. H.. 1972; Experiments in Molecular Genetics pp352–355 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  19. Murillo M., Foglia R., Diller A., Lee S., Leustek T.. 1995; Serine acetyltransferase from Arabidopsis thaliana can functionally complement the cysteine requirement of a cysE mutant strain of Escherichia coli . Cell Mol Biol Res41:425–433
    [Google Scholar]
  20. Nielsen H., Engelbrecht J., Brunak S., von Heijne G.. 1997; Identification of prokaryotic and eukaryotic signal peptides and prediction of the cleavage sites. Protein Eng10:1–6[CrossRef]
    [Google Scholar]
  21. Parker G. F., Higgins T. P., Hawkes T., Robson R. L.. 1999; Rhizobium ( Sinorhizobium ) meliloti phn genes: characterisation and identification of their protein products. J Bacteriol181:389–395
    [Google Scholar]
  22. Patriarca E. J., Riccio A., Taté R., Colonna-Romano S., Iaccarino M., Defez R.. 1993; The ntrBC genes of Rhizobium leguminosarum are part of a complex operon subject to negative regulation. Mol Microbiol9:569–577[CrossRef]
    [Google Scholar]
  23. Poole P. S., Blyth A., Reid C., Walters K.. 1994a; myo -Inositol catabolism and catabolite regulation in Rhizobium leguminosarum bv. viciae. Microbiology140:2787–2795[CrossRef]
    [Google Scholar]
  24. Poole P. S., Schofield N. A., Reid C. J., Drew E. M., Walshaw D. L.. 1994b; Identification of chromosomal genes located downstream of dctD that affect the requirement for calcium and the lipopolysaccharide layer of Rhizobium leguminosarum . Microbiology140:2797–2809[CrossRef]
    [Google Scholar]
  25. Saitou N., Nei M.. 1987; The neighbour-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol4:406–425
    [Google Scholar]
  26. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  27. Studier F. W., Moffatt B. W.. 1986; Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes. J Mol Biol189:113–130[CrossRef]
    [Google Scholar]
  28. Tabor S., Richardson C. C.. 1985; A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci USA82:1074–1078[CrossRef]
    [Google Scholar]
  29. Taté R., Riccio A., Caputo E., Iaccarino M., Patriarca J.. 1999; The Rhizobium etli metZ gene is essential for methionine biosynthesis and nodulation of Phaseolus vulgaris . Mol Plant–Microbe Interact12:24–34[CrossRef]
    [Google Scholar]
  30. Thompson J. D., Higgins D. G., Gibson T. J.. 1994; clustal w: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res22:4673–4680[CrossRef]
    [Google Scholar]
  31. Trinick M. J., Dilworth M. J., Grounds M.. 1976; Factors affecting the reduction of acetylene by root nodules of Lupinus species. New Phytol77:359–370[CrossRef]
    [Google Scholar]
  32. Walshaw D. L., Poole P. S.. 1996; The general l-amino acid permease of Rhizobium leguminosarum is an ABC uptake system that influences efflux of solutes. Mol Microbiol21:1239–1252[CrossRef]
    [Google Scholar]
  33. Walshaw D. L., Reid C. J., Poole P. S.. 1997; The general amino acid permease of Rhizobium leguminosarum is negatively regulated by the Ntr system. FEMS Microbiol Lett152:57–64[CrossRef]
    [Google Scholar]
  34. Zheng L., Cash V. L., Flint D. H., Dean D. R.. 1998; Assembly of iron-sulfur clusters. Identification of an iscSUA-hscBA-fdx gene cluster from Azotobacter vinelandii. J Biol Chem273:13264–13272[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-147-9-2553
Loading
/content/journal/micro/10.1099/00221287-147-9-2553
Loading

Data & Media loading...

Most cited this month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error