RcsA, an unstable positive regulator of capsular polysaccharide synthesis.
RcsA is an unstable positive regulator required for the synthesis of colanic acid capsular polysaccharide in Escherichia coli. Degradation of the RcsA protein in vivo depends on the ATP-dependent Lon protease. DNA sequence analysis of the rcsA gene reveals a single open reading frame for a 23,500-Da highly basic protein (pI = 9.9), consistent with the observed size of the purified subunit of RcsA. The DNA and protein sequences are highly homologous to the rcsA gene and protein from Klebsiella pneumoniae and other species. The carboxy-terminal region of RcsA contains a possible helix-turn-helix DNA-binding motif that resembles sequences found at the carboxy terminus of RcsB, another positive regulator of capsule synthesis, and in several other transcriptional regulators including members of the LuxR family. rcsA62, a mutation in rcsA that leads to increased capsule synthesis, encodes a protein designated RcsA*, which differs from wild-type RcsA only in the replacement of Met-145 by valine. The RcsA* protein is subject to Lon-dependent degradation. The stability of wild-type RcsA in vivo is increased by multicopy RcsB. Conversely, RcsA is degraded more rapidly in rcsB mutant hosts than in wild-type hosts. These results suggest that RcsA and RcsB interact in vivo and are consistent with genetic experiments that indicate an interaction between RcsA and RcsB. Based on these experiments, we propose a model for capsule regulation in which RcsA interacts directly with RcsB to promote transcription of the genes for capsule synthesis.
Stout
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V
Laboratory of Molecular Biology, National Cancer Institute, Bethesda, Maryland 20892.
Torres-Cabassa
A
A
Maurizi
M R
MR
Gutnick
D
D
Gottesman
S
S
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D002872:Chromosome Deletion; D003090:Coliphages / Q000235:genetics; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism; D029968:Escherichia coli Proteins; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D010957:Plasmids; D011135:Polysaccharides, Bacterial / Q000096:biosynthesis*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D012689:Sequence Homology, Nucleic Acid
D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; D011135:Polysaccharides, Bacterial; D011993:Recombinant Fusion Proteins; C067572:RcsA protein, E coli