The Journal of Experimental Medicine
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© The Rockefeller University Press, 0022-1007/1998/12/2047/ $5.00
The Journal of Experimental Medicine, Volume 188, Number 11, December 7, 1998 2047-2056

Articles

Characterization of the galU Gene of Streptococcus pneumoniae Encoding a Uridine Diphosphoglucose Pyrophosphorylase: A Gene Essential for Capsular Polysaccharide Biosynthesis

Marta Mollerach*, Rubens López{ddagger}, and Ernesto García{ddagger}

From the * Departamento de Microbiología, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, 1113 Buenos Aires, Argentina; and the {ddagger} Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28006 Madrid, Spain

The galU gene of Streptococcus pneumoniae has been cloned and sequenced. Escherichia coli cells harboring the recombinant plasmid pMMG2 (galU) overproduced a protein that has been shown to correspond to a uridine 5'-triphosphate:glucose-1-phosphate uridylyltransferase (uridine diphosphoglucose [UDP-Glc] pyrophosphorylase) responsible for the synthesis of UDP-Glc, a key compound in the biosynthesis of polysaccharides. A gene very similar to the S. pneumoniae galU has been found in a partial nucleotide sequence of the Streptococcus pyogenes genome. Knockout galU mutants of type 1 pneumococci are unable to synthesize a detectable capsule. An identical result was found in type 3 S. pneumoniae cells in spite of the fact that these bacteria contain a type-specific gene (cap3C) that also encodes a UDP-Glc pyrophosphorylase. Since eukaryotic UDP-Glc pyrophosphorylases appear to be completely unrelated to their prokaryotic counterparts, we postulate that GalU may be an appropriate target for the search of new drugs to control the pathogenicity of bacteria like pneumococcus and S. pyogenes.

Key Words: pneumococcus • capsule • UDP-Glc pyrophosphorylase • virulence • galU

Address correspondence to Rubens López, Departamento de Microbiología Molecular, Centro de Investigaciones Biológicas, Velázquez 144, 28006 Madrid, Spain. Phone: 34-91-561-1800; Fax: 34-91-562-7518; E-mail: cibl220{at}fresno.csic.es

1 Abbreviations used in this paper: Gal, galactose; GAS, group A streptococci; Glc, glucose; Ln, lincomycin; UDP-GalA, uridine diphosphogalacturonic acid; UDP-Glc, uridine diphosphoglucose; UDP-GlcA, uridine diphosphoglucuronic acid; UDPG:PP, uridine diphosphoglucose pyrophosphorylase.


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