title: Regulation of expression of pectate lyase genes pelA, pelD, and pelE in Erwinia chrysanthemi.
abstract: The regulation of pelA, pelD, and pelE genes encoding three of the five major pectate lyase isoenzymes (PLa, PLd, and PLe) in Erwinia chrysanthemi B374 was analyzed by using genetic fusions to lacZ. These three genes are clustered on a 5-kilobase DNA fragment in the order pelD-pelE-pelA and constitute three independent transcriptional units. We localized the pelDEA cluster near the pro-1 marker on the genetic map of B374 by chromosomal mobilization with RP4::mini-Mu plasmid pULB110. Three classes of regulatory mutations responsible for constitutive pectate lyase synthesis have been described (kdgR, gpiR, and cri). We studied the effects of each mutation on pelE, pelD, and pelA expression independently. The mutations kdgR and gpiR mainly affect the expression of pelE and pelD, although PLa synthesis is slightly increased. The cri mutation results in a low level of constitutive expression of the three pel genes, but it is a pleiotropic mutation since other genes not involved in pectinolysis are also affected. In addition, we demonstrated that exuR, a negative regulatory gene governing the catabolism of hexuronates, does not modify the expression of pel genes. The frequency of gpiR or cri mutations (about 10(-8)) and the resulting constitutivity of pectate lyase synthesis suggest that these genes act as negative regulatory genes in addition to kdgR, which is already known to encode a repressor. Moreover, we found that expression of pel-lac fusions carried on pBR322 derivatives was higher in E. chrysanthemi than in Escherichia coli; this fact suggests the existence of positive regulation of pectate lyase synthesis in E. chrysanthemi.
mesh_terms: D002874:Chromosome Mapping; D003001:Cloning, Molecular; D004885:Erwinia / Q000201:enzymology / Q000235:genetics*; D004926:Escherichia coli; D005786:Gene Expression Regulation; D005798:Genes, Bacterial; D005947:Glucose / Q000378:metabolism; D005990:Glycerol / Q000378:metabolism; D010957:Plasmids; D011133:Polysaccharide-Lyases / Q000235:genetics*; D011401:Promoter Regions, Genetic; D011993:Recombinant Fusion Proteins / Q000235:genetics; D012097:Repressor Proteins / Q000235:genetics; D001616:beta-Galactosidase / Q000235:genetics
chemical_list: D011993:Recombinant Fusion Proteins; D012097:Repressor Proteins; D001616:beta-Galactosidase; D011133:Polysaccharide-Lyases; C023911:pectate lyase; D005947:Glucose; D005990:Glycerol