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Volumn 178, Issue 11, 1996, Pages 3168-3176

The Agrobacterium tumefaciens VirB7 lipoprotein is required for stabilization of VirB proteins during assembly of the T-complex transport apparatus

Author keywords

[No Author keywords available]

Indexed keywords

GENE PRODUCT; LIPOPROTEIN;

EID: 0029891713     PISSN: 00219193     EISSN: None     Source Type: Journal    
DOI: 10.1128/jb.178.11.3168-3176.1996     Document Type: Article
Times cited : (104)

References (51)
  • 1
    • 0026496235 scopus 로고
    • MxiJ, a lipoprotein involved in secretion of Shigella Ipa invasins, is homologous to YscJ, a secretion factor of the Yersinia Yop proteins
    • Allaoui, A., P. J. Sansonetti, and C. Parsot. 1992. MxiJ, a lipoprotein involved in secretion of Shigella Ipa invasins, is homologous to YscJ, a secretion factor of the Yersinia Yop proteins. J. Bacteriol. 174:7661-7669.
    • (1992) J. Bacteriol. , vol.174 , pp. 7661-7669
    • Allaoui, A.1    Sansonetti, P.J.2    Parsot, C.3
  • 2
    • 0021042016 scopus 로고
    • Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors
    • Bagdasarian, M. M., E. Amann, R. Lurz, B. Ruckert, and M. Bagdasarian. 1983. Activity of the hybrid trp-lac (tac) promoter of Escherichia coli in Pseudomonas putida. Construction of broad-host-range, controlled-expression vectors. Gene 26:273-282.
    • (1983) Gene , vol.26 , pp. 273-282
    • Bagdasarian, M.M.1    Amann, E.2    Lurz, R.3    Ruckert, B.4    Bagdasarian, M.5
  • 4
    • 0028126694 scopus 로고
    • Localization and topology of VirB proteins of Agrobacterium tumefaciens
    • Begersbergen, A., S. J. Smith, and P. J. J. Hooykaas. 1994. Localization and topology of VirB proteins of Agrobacterium tumefaciens. Plasmid 32:212-218.
    • (1994) Plasmid , vol.32 , pp. 212-218
    • Begersbergen, A.1    Smith, S.J.2    Hooykaas, P.J.J.3
  • 5
    • 0027500610 scopus 로고
    • The Agrobacterium tumefaciens virB4 gene product is an essential virulence protein requiring an intact nucleoside triphosphate-binding domain
    • Berger, B. R., and P. J. Christie. 1993. The Agrobacterium tumefaciens virB4 gene product is an essential virulence protein requiring an intact nucleoside triphosphate-binding domain. J. Bacteriol. 175:1723-1734.
    • (1993) J. Bacteriol. , vol.175 , pp. 1723-1734
    • Berger, B.R.1    Christie, P.J.2
  • 6
    • 0028243807 scopus 로고
    • Genetic complementation analysis of the Agrobacterium tumefaciens virB operon: VirB2 through virB11 are essential virulence genes
    • Berger, B. R., and P. J. Christie. 1994. Genetic complementation analysis of the Agrobacterium tumefaciens virB operon: virB2 through virB11 are essential virulence genes. J. Bacteriol. 176:3646-3660.
    • (1994) J. Bacteriol. , vol.176 , pp. 3646-3660
    • Berger, B.R.1    Christie, P.J.2
  • 7
    • 0026099076 scopus 로고
    • Controlled expression of the transcriptional activator gene virG in Agrobacterium tumefaciens by using the Escherichia coli lac promoter
    • Chen, C.-Y., and S. C. Winans. 1991. Controlled expression of the transcriptional activator gene virG in Agrobacterium tumefaciens by using the Escherichia coli lac promoter. J. Bacteriol. 173:1139-1144.
    • (1991) J. Bacteriol. , vol.173 , pp. 1139-1144
    • Chen, C.-Y.1    Winans, S.C.2
  • 8
    • 0022999829 scopus 로고
    • Trimeric structure and localization of the major lipoprotein in the cell surface of Escherichia coli
    • Choi, D.-S., H. Yamada, T. Mizumo, and S. Mizushima. 1986. Trimeric structure and localization of the major lipoprotein in the cell surface of Escherichia coli. J. Biol. Chem. 261:8953-8957.
    • (1986) J. Biol. Chem. , vol.261 , pp. 8953-8957
    • Choi, D.-S.1    Yamada, H.2    Mizumo, T.3    Mizushima, S.4
  • 9
    • 0024805123 scopus 로고
    • A gene required for transfer of T-DNA to plants encodes an ATPase with autophosphorylating activity
    • Christie, P. J., J. E. Ward, M. P. Gordon, and E. W. Nester. 1989. A gene required for transfer of T-DNA to plants encodes an ATPase with autophosphorylating activity. Proc. Natl. Acad. Sci. USA 86:9677-9681.
    • (1989) Proc. Natl. Acad. Sci. USA , vol.86 , pp. 9677-9681
    • Christie, P.J.1    Ward, J.E.2    Gordon, M.P.3    Nester, E.W.4
  • 10
    • 0024022150 scopus 로고
    • The Agrobacterium tumefaciens virE2 gene product is a single-stranded-DNA-binding protein that associates with T-DNA
    • Christie, P. J., J. E. Ward, S. C. Winans, and E. W. Nester. 1988. The Agrobacterium tumefaciens virE2 gene product is a single-stranded-DNA-binding protein that associates with T-DNA. J. Bacteriol. 170:2659-2667.
    • (1988) J. Bacteriol. , vol.170 , pp. 2659-2667
    • Christie, P.J.1    Ward, J.E.2    Winans, S.C.3    Nester, E.W.4
  • 11
    • 0027455152 scopus 로고
    • Transport of nucleic acids through membrane channels: Snaking through small holes
    • Citovsky, V., and P. Zambryski. 1993. Transport of nucleic acids through membrane channels: snaking through small holes. Annu. Rev. Microbiol. 47:167-197.
    • (1993) Annu. Rev. Microbiol. , vol.47 , pp. 167-197
    • Citovsky, V.1    Zambryski, P.2
  • 13
    • 0023441713 scopus 로고
    • Cloning and expression in Escherichia coli of the Klebsiella pneumoniae genes for production, surface localization and secretion of the lipoprotein pullulanase
    • de'Enfert, C., A. Ryter, and A. P. Pugsley. 1987. Cloning and expression in Escherichia coli of the Klebsiella pneumoniae genes for production, surface localization and secretion of the lipoprotein pullulanase. EMBO J. 6:3531-3538.
    • (1987) EMBO J. , vol.6 , pp. 3531-3538
    • De'Enfert, C.1    Ryter, A.2    Pugsley, A.P.3
  • 14
    • 0028260496 scopus 로고
    • Molecular analysis of the F plasmid traVR region: TraV encodes a lipoprotein
    • Doran, T. J., S. M. Loh, N. Firth, and R. A. Skurray. 1994. Molecular analysis of the F plasmid traVR region: traV encodes a lipoprotein. J. Bacteriol. 176:4182-4186.
    • (1994) J. Bacteriol. , vol.176 , pp. 4182-4186
    • Doran, T.J.1    Loh, S.M.2    Firth, N.3    Skurray, R.A.4
  • 15
    • 0029941238 scopus 로고    scopus 로고
    • The Agrobacterium tumefaciens virB7 gene product, a proposed component of the T-complex transport apparatus, is a membrane-associated lipoprotein exposed at the periplasmic surface
    • Fernandez, D., T. A. T. Dang, G. M. Spudich, X.-R. Zhou, B. R. Berger, and P. J. Christie. 1996. The Agrobacterium tumefaciens virB7 gene product, a proposed component of the T-complex transport apparatus, is a membrane-associated lipoprotein exposed at the periplasmic surface. J. Bacteriol. 178: 3156-3167.
    • (1996) J. Bacteriol. , vol.178 , pp. 3156-3167
    • Fernandez, D.1    Dang, T.A.T.2    Spudich, G.M.3    Zhou, X.-R.4    Berger, B.R.5    Christie, P.J.6
  • 16
    • 0029127480 scopus 로고
    • Interactions of VirB9, -10, and -11 with the membrane fraction of Agrobacterium tumefaciens; solubility studies provide evidence for tight associations
    • Finberg, K. E., T. R. Muth, S. P. Young, J. B. Maken, S. M. Heitritter, A. N. Binns, and L. M. Banta. 1995. Interactions of VirB9, -10, and -11 with the membrane fraction of Agrobacterium tumefaciens; solubility studies provide evidence for tight associations. J. Bacteriol. 177:4881-4889.
    • (1995) J. Bacteriol. , vol.177 , pp. 4881-4889
    • Finberg, K.E.1    Muth, T.R.2    Young, S.P.3    Maken, J.B.4    Heitritter, S.M.5    Binns, A.N.6    Banta, L.M.7
  • 17
    • 0028569497 scopus 로고
    • An essential virulence protein of Agrobacterium tumefaciens, VirB4, requires an intact mononucleotide binding domain to function in transfer of T-DNA
    • Fullner, K. J., K. M. Stephens, and E. W. Nester. 1994. An essential virulence protein of Agrobacterium tumefaciens, VirB4, requires an intact mononucleotide binding domain to function in transfer of T-DNA. Mol. Gen. Genet. 245:704-715.
    • (1994) Mol. Gen. Genet. , vol.245 , pp. 704-715
    • Fullner, K.J.1    Stephens, K.M.2    Nester, E.W.3
  • 18
    • 0019847082 scopus 로고
    • Genetic analysis of crown gall: Fine structure map of the T-DNA by site-directed mutagenesis
    • Garfinkel, D. J., R. B. Simpson, L. W. Ream, F. F. White, M. P. Gordon, and E. W. Nester. 1981. Genetic analysis of crown gall: fine structure map of the T-DNA by site-directed mutagenesis. Cell 27:143-153.
    • (1981) Cell , vol.27 , pp. 143-153
    • Garfinkel, D.J.1    Simpson, R.B.2    Ream, L.W.3    White, F.F.4    Gordon, M.P.5    Nester, E.W.6
  • 19
    • 0029148738 scopus 로고
    • Bacterial conjugation mediated by plasmid RP4: RSF1010 mobilization, donor-specific phage production, and pilus production require the same Tra2 core components of a proposed DNA transport complex
    • Haase, J., R. Lurz, A. M. Grahn, D. H. Bamford, and E. Lanka. 1995. Bacterial conjugation mediated by plasmid RP4: RSF1010 mobilization, donor-specific phage production, and pilus production require the same Tra2 core components of a proposed DNA transport complex. J. Bacteriol. 177:4779-4791.
    • (1995) J. Bacteriol. , vol.177 , pp. 4779-4791
    • Haase, J.1    Lurz, R.2    Grahn, A.M.3    Bamford, D.H.4    Lanka, E.5
  • 21
    • 0023239517 scopus 로고
    • Functional inactivation of genes by dominant negative mutations
    • Herskowitz, I. 1987. Functional inactivation of genes by dominant negative mutations. Nature (London) 329:219-222.
    • (1987) Nature (London) , vol.329 , pp. 219-222
    • Herskowitz, I.1
  • 22
    • 0028085879 scopus 로고
    • The product of the virB4 gene of Agrobacterium tumefaciens promotes accumulation of VirB3 protein
    • Jones, A. L., D. Shirasu, and C. I. Kado. 1994. The product of the virB4 gene of Agrobacterium tumefaciens promotes accumulation of VirB3 protein. J. Bacteriol. 176:5255-5261.
    • (1994) J. Bacteriol. , vol.176 , pp. 5255-5261
    • Jones, A.L.1    Shirasu, D.2    Kado, C.I.3
  • 23
    • 0028200971 scopus 로고
    • Promiscuous DNA transfer system of Agrobacterium tumefaciens: Role of the virB operon in sex pilus assembly and synthesis
    • Kado, C. I. 1994. Promiscuous DNA transfer system of Agrobacterium tumefaciens: role of the virB operon in sex pilus assembly and synthesis. Mol. Microbiol. 12:17-22.
    • (1994) Mol. Microbiol. , vol.12 , pp. 17-22
    • Kado, C.I.1
  • 24
    • 0025355447 scopus 로고
    • The virB operon of Agrobacterium tumefaciens pTiC58 encodes 11 open reading frames
    • Kuldau, G. A., G. DeVos, J. Owen, G. McCaffrey, and P. Zambryski. 1990. The virB operon of Agrobacterium tumefaciens pTiC58 encodes 11 open reading frames. Mol. Gen. Genet. 221:256-266.
    • (1990) Mol. Gen. Genet. , vol.221 , pp. 256-266
    • Kuldau, G.A.1    DeVos, G.2    Owen, J.3    McCaffrey, G.4    Zambryski, P.5
  • 25
    • 0026730667 scopus 로고
    • Sequence similarities between the RP4 Tra2 and the Ti VirB region strongly support the conjugation model for T-DNA transfer
    • Lessl, M., D. Balzer, W. Pansegrau, and E. Lanka. 1992. Sequence similarities between the RP4 Tra2 and the Ti VirB region strongly support the conjugation model for T-DNA transfer. J. Biol. Chem. 267:20471-20480.
    • (1992) J. Biol. Chem. , vol.267 , pp. 20471-20480
    • Lessl, M.1    Balzer, D.2    Pansegrau, W.3    Lanka, E.4
  • 26
    • 0028315917 scopus 로고
    • Common mechanisms in bacterial conjugation and Ti-mediated T-DNA transfer to plant cells
    • Lessl, M., and E. Lanka. 1994. Common mechanisms in bacterial conjugation and Ti-mediated T-DNA transfer to plant cells. Cell 77:321-324.
    • (1994) Cell , vol.77 , pp. 321-324
    • Lessl, M.1    Lanka, E.2
  • 27
    • 0028981022 scopus 로고
    • A novel periplasmic carrier protein involved in the sorting and transport of Escherichia coli lipoproteins destined for the outer membrane
    • Matsuyama, S., T. Tajima, and H. Tokuda. 1995. A novel periplasmic carrier protein involved in the sorting and transport of Escherichia coli lipoproteins destined for the outer membrane. EMBO J. 14:3365-3372.
    • (1995) EMBO J. , vol.14 , pp. 3365-3372
    • Matsuyama, S.1    Tajima, T.2    Tokuda, H.3
  • 29
  • 30
    • 0021328518 scopus 로고
    • The product of the F sex factor traT surface exclusion gene is a lipoprotein
    • Perumal, N. B., and E. G. Micnkley, Jr. 1984. The product of the F sex factor traT surface exclusion gene is a lipoprotein. J. Biol. Chem. 259:5357-5360.
    • (1984) J. Biol. Chem. , vol.259 , pp. 5357-5360
    • Perumal, N.B.1    Micnkley Jr., E.G.2
  • 31
    • 0027985773 scopus 로고
    • Common ancestry between IncN conjugal transfer genes and macromolecular export systems of plant and animal pathogens
    • Pohlman, R. F., H. D. Genetti, and S. C. Winans. 1994. Common ancestry between IncN conjugal transfer genes and macromolecular export systems of plant and animal pathogens. Mol. Microbiol. 14:655-668.
    • (1994) Mol. Microbiol. , vol.14 , pp. 655-668
    • Pohlman, R.F.1    Genetti, H.D.2    Winans, S.C.3
  • 32
    • 0027450561 scopus 로고
    • The complete general secretory pathway in gram-negative bacteria
    • Pugsley, A. P. 1993. The complete general secretory pathway in gram-negative bacteria. Microbiol. Rev. 57:50-108.
    • (1993) Microbiol. Rev. , vol.57 , pp. 50-108
    • Pugsley, A.P.1
  • 34
    • 0028318253 scopus 로고
    • SecF stabilizes SecD and SecY, components of the protein translocation machinery of the Escherichia coli cytoplasmic membrane
    • Sagara, K., S.-I. Matsuyama, and S. Mizushima. 1994. SecF stabilizes SecD and SecY, components of the protein translocation machinery of the Escherichia coli cytoplasmic membrane. J. Bacteriol. 176:4111-4116.
    • (1994) J. Bacteriol. , vol.176 , pp. 4111-4116
    • Sagara, K.1    Matsuyama, S.-I.2    Mizushima, S.3
  • 35
    • 0023472472 scopus 로고
    • Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa
    • Schagger, H., and G. von Jagow. 1987. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166:368-379.
    • (1987) Anal. Biochem. , vol.166 , pp. 368-379
    • Schagger, H.1    Von Jagow, G.2
  • 36
    • 0026573593 scopus 로고
    • SecY variants that interfere with Escherichia colt protein export in the presence of normal secY
    • Shimoike, T., Y. Akiyama, T. Baba, T. Taura, and K. Ho. 1992. SecY variants that interfere with Escherichia colt protein export in the presence of normal secY. Mol. Microbiol. 6:1205-1210.
    • (1992) Mol. Microbiol. , vol.6 , pp. 1205-1210
    • Shimoike, T.1    Akiyama, Y.2    Baba, T.3    Taura, T.4    Ho, K.5
  • 37
    • 0027291335 scopus 로고
    • Membrane location of the Ti plasmid VirB proteins involved in the biosynthesis of a pilin-like conjugative structure on Agrobacterium tumefaciens
    • Shirasu, K., and C. I. Kado. 1993. Membrane location of the Ti plasmid VirB proteins involved in the biosynthesis of a pilin-like conjugative structure on Agrobacterium tumefaciens. FEMS Microbiol. Lett. 111:287-294.
    • (1993) FEMS Microbiol. Lett. , vol.111 , pp. 287-294
    • Shirasu, K.1    Kado, C.I.2
  • 38
    • 0025122756 scopus 로고
    • Characterization of the virB operon of an Agrobacterium tumefaciens Ti plasmid: Nucleotide sequence and protein analysis
    • Shirasu, K., P. Morel, and C. I. Kado. 1990. Characterization of the virB operon of an Agrobacterium tumefaciens Ti plasmid: nucleotide sequence and protein analysis. Mol. Microbiol. 4:1153-1163.
    • (1990) Mol. Microbiol. , vol.4 , pp. 1153-1163
    • Shirasu, K.1    Morel, P.2    Kado, C.I.3
  • 39
    • 85035165958 scopus 로고    scopus 로고
    • Intermolecular disulfide bonds stabilize VirB7 homodimers and VirB7/VirB9 heterodimers during biogenesis of the Agrobacterium tumefaciens T-complex transport apparatus
    • in press
    • Spudich, G. M., D. Fernandez, X.-R. Zhoti, and P. J. Christie. Intermolecular disulfide bonds stabilize VirB7 homodimers and VirB7/VirB9 heterodimers during biogenesis of the Agrobacterium tumefaciens T-complex transport apparatus. Proc. Natl. Acad. Sci. USA, in press.
    • Proc. Natl. Acad. Sci. USA
    • Spudich, G.M.1    Fernandez, D.2    Zhoti, X.-R.3    Christie, P.J.4
  • 40
    • 0022744866 scopus 로고
    • The genetic and transcriptional organization of the vir region of the A6 Ti-plasmid at Agrobacterium tumefaciens
    • Stachel, S. E., and E. W. Nester. 1986. The genetic and transcriptional organization of the vir region of the A6 Ti-plasmid at Agrobacterium tumefaciens. EMBO J. 5:1445-1454.
    • (1986) EMBO J. , vol.5 , pp. 1445-1454
    • Stachel, S.E.1    Nester, E.W.2
  • 41
    • 0028842065 scopus 로고
    • Agrobacterium tumefaciens VirB11 protein requires a consensus nucleotide binding site for function in virulence
    • Stephens, K. M., C. Roush, and E. Nester. 1995. Agrobacterium tumefaciens VirB11 protein requires a consensus nucleotide binding site for function in virulence. J. Bacteriol. 177:27-36.
    • (1995) J. Bacteriol. , vol.177 , pp. 27-36
    • Stephens, K.M.1    Roush, C.2    Nester, E.3
  • 43
    • 0027186527 scopus 로고
    • Subcellular localization of seven VirB proteins of Agrobacterium tumefaciens: Implications for the formation of a T-DNA transport structure
    • Thorstenson, Y. R., G. A. Kuldau, and P. C. Zambryski. 1993. Subcellular localization of seven VirB proteins of Agrobacterium tumefaciens: implications for the formation of a T-DNA transport structure. J. Bacteriol. 175: 5233-5241.
    • (1993) J. Bacteriol. , vol.175 , pp. 5233-5241
    • Thorstenson, Y.R.1    Kuldau, G.A.2    Zambryski, P.C.3
  • 44
    • 0028262147 scopus 로고
    • The essential virulence protein VirB8 localizes to the inner membrane of Agrobacterium tumefaciens
    • Thorstenson, Y. R., and P. C. Zambryski. 1994. The essential virulence protein VirB8 localizes to the inner membrane of Agrobacterium tumefaciens. J. Bacteriol. 176:1711-1717.
    • (1994) J. Bacteriol. , vol.176 , pp. 1711-1717
    • Thorstenson, Y.R.1    Zambryski, P.C.2
  • 45
    • 0023931033 scopus 로고
    • Characterization of the virB operon from an Agrobacterium tumefaciens Ti plasmid
    • Ward, J. E., D. E. Akiyoshi, D. Regier, A. Datta, M. P. Gordon, and E. W. Nester. 1988. Characterization of the virB operon from an Agrobacterium tumefaciens Ti plasmid. J. Biol. Chem. 263:5804-5814.
    • (1988) J. Biol. Chem. , vol.263 , pp. 5804-5814
    • Ward, J.E.1    Akiyoshi, D.E.2    Regier, D.3    Datta, A.4    Gordon, M.P.5    Nester, E.W.6
  • 47
    • 0025108802 scopus 로고
    • Complementation analysis of Agrobacterium tumefaciens Ti plasmid virB genes by use of a vir promoter expression vector: VirB9, virB10, and virB11 are essential virulence genes
    • Ward, J. E., Jr., E. M. Dale, P. J. Christie, E. W. Nester, and A. N. Binns. 1990. Complementation analysis of Agrobacterium tumefaciens Ti plasmid virB genes by use of a vir promoter expression vector: virB9, virB10, and virB11 are essential virulence genes. J. Bacteriol. 172:5187-5199.
    • (1990) J. Bacteriol. , vol.172 , pp. 5187-5199
    • Ward Jr., J.E.1    Dale, E.M.2    Christie, P.J.3    Nester, E.W.4    Binns, A.N.5
  • 48
    • 0025181295 scopus 로고
    • Identification of a VirB10 protein aggregate in the inner membrane of Agrobacterium tumefaciens
    • Ward, J. E., Jr., E. M. Dale, E. W. Nester, and A. N. Binns. 1990. Identification of a VirB10 protein aggregate in the inner membrane of Agrobacterium tumefaciens. J. Bacteriol. 172:5200-5210.
    • (1990) J. Bacteriol. , vol.172 , pp. 5200-5210
    • Ward Jr., J.E.1    Dale, E.M.2    Nester, E.W.3    Binns, A.N.4
  • 49
    • 0027518747 scopus 로고
    • Molecular characterization of an operon required for pertussis toxin secretion
    • Weiss, A. A., F. D. Johnson, and D. L. Burns. 1993. Molecular characterization of an operon required for pertussis toxin secretion. Proc. Natl. Acad. Sci. USA 90:297-2974.
    • (1993) Proc. Natl. Acad. Sci. USA , vol.90 , pp. 297-2974
    • Weiss, A.A.1    Johnson, F.D.2    Burns, D.L.3
  • 50
    • 0029968272 scopus 로고    scopus 로고
    • Adaptation of a conjugal transfer system for the export of pathogenic macromolecules
    • Winans, S. C., D. L. Burns, and P. J. Christie. 1996. Adaptation of a conjugal transfer system for the export of pathogenic macromolecules. Trends Microbiol. 4:1616-1622.
    • (1996) Trends Microbiol. , vol.4 , pp. 1616-1622
    • Winans, S.C.1    Burns, D.L.2    Christie, P.J.3
  • 51
    • 0000398313 scopus 로고
    • Chronicles from the Agrobacterium-plant cell DNA transfer story
    • Zambryski, P. C. 1992. Chronicles from the Agrobacterium-plant cell DNA transfer story. Annu. Rev. Plant Physiol. Plant Mol. Biol. 43:465-490.
    • (1992) Annu. Rev. Plant Physiol. Plant Mol. Biol. , vol.43 , pp. 465-490
    • Zambryski, P.C.1


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