Identification and molecular cloning of a unique hyaluronan synthase from Pasteurella multocida

Type A Pasteurella multocida , a prevalent animal pathogen, employs a hyaluronan [HA] polysaccharide capsule to avoid host defenses. We utilized transposon insertional mutagenesis to identify the P. multocida HA synthase, the enzyme that polymerizes HA. A DNA fragment from a wild-type genomic librar...

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Bibliographic Details
Published in:	The Journal of biological chemistry Vol. 273; no. 14; pp. 8454 - 8458
Main Authors:	DeAngelis, P.L. (University of Oklahoma Health Sciences Center, Oklahoma City.), Jing, W, Drake, R.R, Achyuthan, A.M
Format:	Journal Article
Language:	English
Published:	United States American Society for Biochemistry and Molecular Biology 03-04-1998
Subjects:	ACIDE HYALURONIQUE ACIDO HIALURONICO Amino Acid Sequence AMINO ACID SEQUENCES Base Sequence BIOCHEMISTRY BIOCHIMIE BIOQUIMICA CARBOHYDRATE METABOLISM CELL MEMBRANES CHEMICAL COMPOSITION CLONACION CLONACION MOLECULAR CLONAGE CLONAGE MOLECULAIRE CLONING Cloning, Molecular COMPOSICION QUIMICA COMPOSITION CHIMIQUE CYTOCHEMISTRY GENBANK/AF036004 Genes, Bacterial GLICOSILTRANSFERASAS Glucuronosyltransferase - genetics GLYCOSYLTRANSFERASE GLYCOSYLTRANSFERASES HEXOSYLTRANSFERASES Hyaluronan Synthases HYALURONIC ACID MEMBRANAS CELULARES MEMBRANE CELLULAIRE Membrane Proteins METABOLISME DES GLUCIDES METABOLISMO DE CARBOHIDRATOS MOLECULAR CLONING MOLECULAR SEQUENCE DATA MUTANT MUTANTES MUTANTS NUCLEOTIDE SEQUENCE OPEN READING FRAMES PASTEURELLA MULTOCIDA Pasteurella multocida - genetics SECUENCIA NUCLEOTIDICA Sequence Alignment SEQUENCE NUCLEOTIDIQUE Transferases Xenopus Proteins
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Summary:	Type A Pasteurella multocida , a prevalent animal pathogen, employs a hyaluronan [HA] polysaccharide capsule to avoid host defenses. We utilized transposon insertional mutagenesis to identify the P. multocida HA synthase, the enzyme that polymerizes HA. A DNA fragment from a wild-type genomic library could direct HA production in vivo in Escherichia coli , a bacterium that normally does not produce HA. Analysis of truncated plasmids derived from the original clone indicated that an open reading frame encoding a 972-residue protein was responsible for HA polymerization. This identification was confirmed by expression cloning in E. coli ; we observed HA capsule formation in vivo and detected activity in membrane preparations in vitro . The polypeptide size was verified by photoaffinity labeling of the native P. multocida HA synthase with azido-UDP sugar analogs. Overall, the P. multocida sequence is not very similar to the other known HA synthases from streptococci, PBCV-1 virus, or vertebrates. Instead, a portion of the central region of the new enzyme is more homologous to the amino termini of other bacterial glycosyltransferases that produce different capsular polysaccharides or lipopolysaccharides. In summary, we have discovered a unique HA synthase that differs in sequence and predicted topology from the other known enzymes.
Bibliography:	1997078743 L73 ObjectType-Article-2 SourceType-Scholarly Journals-1 ObjectType-Feature-1 content type line 23
ISSN:	0021-9258 1083-351X
DOI:	10.1074/jbc.273.14.8454