Role for [corrected] Agrobacterium tumefaciens ChvA protein in export of beta-1,2-glucan.
Functional chvA and chvB genes are required for attachment of Agrobacterium tumefaciens to plant cells, an early step in crown gall tumor formation. Strains defective in these loci do not secrete normal amounts of cyclic beta-1,2-glucan. Whereas chvB is required for beta-1,2-glucan synthesis, the role of chvA in glucan synthesis or export has not been clearly defined. We found that cultures of chvA mutants contained as much neutral beta-1,2-glucan in the cell pellets as did the wild type, with no detectable accumulation of glucan in the culture supernatant. The cytoplasm of chvA mutant cells contained over three times more soluble beta-1,2-glucan than did the cytoplasm of the wild-type parent. Unlike the wild type, chvA mutants contained no detectable periplasmic glucan. The amino acid sequence of chvA is highly homologous to the sequences of bacterial and eucaryotic export proteins, as observed previously in the case of ndvA, a rhizobial homolog of chvA. Strong sequence homology within this family of export proteins is concentrated in the carboxy-terminal portions of the proteins, but placement of consensus ATP-binding sites, internal signal sequences, and hydrophobic domains are conserved over their entire lengths. These data suggest a model for beta-1,2-glucan synthesis in A. tumefaciens in which glucan is synthesized inside the inner membrane with the participation of ChvB and transported across the inner membrane with the participation of ChvA.
Cangelosi
G A
GA
Department of Microbiology, University of Washington, Seattle 98195.
Martinetti
G
G
Leigh
J A
JA
Lee
C C
CC
Thienes
C
C
Theines
C
C
Nester
E W
EW
D018528:ATP-Binding Cassette Transporters; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000502:physiology; D001483:Base Sequence; D005796:Genes; D005798:Genes, Bacterial; D005936:Glucans / Q000096:biosynthesis* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D009154:Mutation; D011487:Protein Conformation; D012231:Rhizobium / Q000235:genetics* / Q000378:metabolism
D018528:ATP-Binding Cassette Transporters; D001426:Bacterial Proteins; C058770:ChvA protein, Agrobacterium tumefaciens; D005936:Glucans