Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene.
The chvB operon of Agrobacterium tumefaciens is required for bacterial attachment to plant cells and for efficient crown gall tumor formation. As defined by the virulence phenotypes of mutants with transposon insertions mapping in the region, the operon was previously mapped to a 5-kilobase (kb) str...
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todo:paper_08940282_v1_n3_p121_Zorreguieta2023-10-03T15:41:54Z Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. Zorreguieta, A. Geremia, R.A. Cavaignac, S. Cangelosi, G.A. Nester, E.W. Ugalde, R.A. beta 1,2 glucan beta galactosidase beta glucan beta-1,2-glucan glucan trypsin article bacterial chromosome bacterial gene biosynthesis genetics high performance liquid chromatography molecular cloning operon pathogenicity phenotype plasmid polyacrylamide gel electrophoresis restriction mapping Rhizobium radiobacter transposon virulence beta-Galactosidase beta-Glucans Chromatography, High Pressure Liquid Chromosomes, Bacterial Cloning, Molecular DNA Transposable Elements Electrophoresis, Polyacrylamide Gel Genes, Bacterial Glucans Operon Phenotype Plasmids Restriction Mapping Rhizobium radiobacter Trypsin Virulence The chvB operon of Agrobacterium tumefaciens is required for bacterial attachment to plant cells and for efficient crown gall tumor formation. As defined by the virulence phenotypes of mutants with transposon insertions mapping in the region, the operon was previously mapped to a 5-kilobase (kb) stretch of chromosomal DNA. We report here that the operon is actually about 8.5 kb long and that it contains a 7-kb gene coding for a large membrane protein involved in the synthesis of cyclic beta-1,2-glucan. Mutants with transposon insertions within the 5-kb phenotypically defined operon do not synthesize this functional protein, do not synthesize beta-1,2-glucan, and do not form tumors. However, mutants with insertions that map up to 3.5 kb downstream of the phenotypically defined operon synthesize truncated proteins that are active in beta-1,2-glucan synthesis. These mutants form tumors. The truncated proteins correspond closely in size with the map positions of the insertions, suggesting that the insertions truncate the proteins by translational termination. A plasmid that contains only the phenotypically defined chvB operon also codes for a truncated protein. A fusion product between the protein and beta-galactosidase carried on a Tn3-HoHo1 insertion was observed in one mutant. Partial trypsin digestion of wild-type inner membranes generated truncated proteins that were active in beta-1,2-glucan synthesis, demonstrating that a large portion of the protein is not required for beta-1,2-glucan synthesis. The correlation between beta-1,2-glucan synthesis by the truncated proteins and tumorigenesis strongly implicates the polysaccharide product of this protein in tumor formation. Fil:Zorreguieta, A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Geremia, R.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Ugalde, R.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_08940282_v1_n3_p121_Zorreguieta |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
beta 1,2 glucan beta galactosidase beta glucan beta-1,2-glucan glucan trypsin article bacterial chromosome bacterial gene biosynthesis genetics high performance liquid chromatography molecular cloning operon pathogenicity phenotype plasmid polyacrylamide gel electrophoresis restriction mapping Rhizobium radiobacter transposon virulence beta-Galactosidase beta-Glucans Chromatography, High Pressure Liquid Chromosomes, Bacterial Cloning, Molecular DNA Transposable Elements Electrophoresis, Polyacrylamide Gel Genes, Bacterial Glucans Operon Phenotype Plasmids Restriction Mapping Rhizobium radiobacter Trypsin Virulence |
| spellingShingle |
beta 1,2 glucan beta galactosidase beta glucan beta-1,2-glucan glucan trypsin article bacterial chromosome bacterial gene biosynthesis genetics high performance liquid chromatography molecular cloning operon pathogenicity phenotype plasmid polyacrylamide gel electrophoresis restriction mapping Rhizobium radiobacter transposon virulence beta-Galactosidase beta-Glucans Chromatography, High Pressure Liquid Chromosomes, Bacterial Cloning, Molecular DNA Transposable Elements Electrophoresis, Polyacrylamide Gel Genes, Bacterial Glucans Operon Phenotype Plasmids Restriction Mapping Rhizobium radiobacter Trypsin Virulence Zorreguieta, A. Geremia, R.A. Cavaignac, S. Cangelosi, G.A. Nester, E.W. Ugalde, R.A. Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. |
| topic_facet |
beta 1,2 glucan beta galactosidase beta glucan beta-1,2-glucan glucan trypsin article bacterial chromosome bacterial gene biosynthesis genetics high performance liquid chromatography molecular cloning operon pathogenicity phenotype plasmid polyacrylamide gel electrophoresis restriction mapping Rhizobium radiobacter transposon virulence beta-Galactosidase beta-Glucans Chromatography, High Pressure Liquid Chromosomes, Bacterial Cloning, Molecular DNA Transposable Elements Electrophoresis, Polyacrylamide Gel Genes, Bacterial Glucans Operon Phenotype Plasmids Restriction Mapping Rhizobium radiobacter Trypsin Virulence |
| description |
The chvB operon of Agrobacterium tumefaciens is required for bacterial attachment to plant cells and for efficient crown gall tumor formation. As defined by the virulence phenotypes of mutants with transposon insertions mapping in the region, the operon was previously mapped to a 5-kilobase (kb) stretch of chromosomal DNA. We report here that the operon is actually about 8.5 kb long and that it contains a 7-kb gene coding for a large membrane protein involved in the synthesis of cyclic beta-1,2-glucan. Mutants with transposon insertions within the 5-kb phenotypically defined operon do not synthesize this functional protein, do not synthesize beta-1,2-glucan, and do not form tumors. However, mutants with insertions that map up to 3.5 kb downstream of the phenotypically defined operon synthesize truncated proteins that are active in beta-1,2-glucan synthesis. These mutants form tumors. The truncated proteins correspond closely in size with the map positions of the insertions, suggesting that the insertions truncate the proteins by translational termination. A plasmid that contains only the phenotypically defined chvB operon also codes for a truncated protein. A fusion product between the protein and beta-galactosidase carried on a Tn3-HoHo1 insertion was observed in one mutant. Partial trypsin digestion of wild-type inner membranes generated truncated proteins that were active in beta-1,2-glucan synthesis, demonstrating that a large portion of the protein is not required for beta-1,2-glucan synthesis. The correlation between beta-1,2-glucan synthesis by the truncated proteins and tumorigenesis strongly implicates the polysaccharide product of this protein in tumor formation. |
| format |
JOUR |
| author |
Zorreguieta, A. Geremia, R.A. Cavaignac, S. Cangelosi, G.A. Nester, E.W. Ugalde, R.A. |
| author_facet |
Zorreguieta, A. Geremia, R.A. Cavaignac, S. Cangelosi, G.A. Nester, E.W. Ugalde, R.A. |
| author_sort |
Zorreguieta, A. |
| title |
Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. |
| title_short |
Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. |
| title_full |
Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. |
| title_fullStr |
Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. |
| title_full_unstemmed |
Identification of the product of an Agrobacterium tumefaciens chromosomal virulence gene. |
| title_sort |
identification of the product of an agrobacterium tumefaciens chromosomal virulence gene. |
| url |
http://hdl.handle.net/20.500.12110/paper_08940282_v1_n3_p121_Zorreguieta |
| work_keys_str_mv |
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| _version_ |
1807324247736975360 |