Mammalian vaults are ribonucleoprotein (RNP) complexes, composed of a small ribonucleic acid and three proteins of 100, 193, and 240 kD in size. The 100-kD major vault protein (MVP) accounts for >70% of the particle mass. We have identified the 193-kD vault protein by its interaction with the MVP in a yeast two-hybrid screen and confirmed its identity by peptide sequence analysis. Analysis of the protein sequence revealed a region of ∼350 amino acids that shares 28% identity with the catalytic domain of poly(ADP-ribose) polymerase (PARP). PARP is a nuclear protein that catalyzes the formation of ADP-ribose polymers in response to DNA damage. The catalytic domain of p193 was expressed and purified from bacterial extracts. Like PARP, this domain is capable of catalyzing a poly(ADP-ribosyl)ation reaction; thus, the 193-kD protein is a new PARP. Purified vaults also contain the poly(ADP-ribosyl)ation activity, indicating that the assembled particle retains enzymatic activity. Furthermore, we show that one substrate for this vault-associated PARP activity is the MVP. Immunofluorescence and biochemical data reveal that p193 protein is not entirely associated with the vault particle, suggesting that it may interact with other protein(s). A portion of p193 is nuclear and localizes to the mitotic spindle.
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6 September 1999
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September 06 1999
The 193-Kd Vault Protein, Vparp, Is a Novel Poly(Adp-Ribose) Polymerase
Valerie A. Kickhoefer,
Valerie A. Kickhoefer
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
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Amara C. Siva,
Amara C. Siva
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
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Nancy L. Kedersha,
Nancy L. Kedersha
bDivision of Rheumatology and Immunology, Brigham and Women's Hospital, Boston, Massachusetts 02115
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Elisabeth M. Inman,
Elisabeth M. Inman
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
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Cristina Ruland,
Cristina Ruland
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
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Michel Streuli,
Michel Streuli
cDepartment of Cancer, Immunology, and AIDS, Dana-Farber Cancer Institute, Boston, Massachusetts 02115
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Leonard H. Rome
Leonard H. Rome
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
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Valerie A. Kickhoefer
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
Amara C. Siva
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
Nancy L. Kedersha
bDivision of Rheumatology and Immunology, Brigham and Women's Hospital, Boston, Massachusetts 02115
Elisabeth M. Inman
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
Cristina Ruland
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
Michel Streuli
cDepartment of Cancer, Immunology, and AIDS, Dana-Farber Cancer Institute, Boston, Massachusetts 02115
Leonard H. Rome
aDepartment of Biological Chemistry, University of California, Los Angeles School of Medicine, Los Angeles, California 90095-1737
1.used in this paper: 3ABA, 3-aminobenzamide; aa, amino acid(s); BRCT, BRCA1 COOH terminus; GST, glutathione S-transferase; MVP, major vault protein; PARP, poly(ADP-ribose) polymerase; RACE, rapid amplification of cDNA ends; TEP1, telomerase-associated protein 1; vRNA, vault-associated ribonucleic acid; VPARP, vault PARP; VSVG, vesicular stomatitis virus glycoprotein
Received:
June 11 1999
Revision Requested:
July 27 1999
Accepted:
August 04 1999
Online ISSN: 1540-8140
Print ISSN: 0021-9525
© 1999 The Rockefeller University Press
1999
The Rockefeller University Press
J Cell Biol (1999) 146 (5): 917–928.
Article history
Received:
June 11 1999
Revision Requested:
July 27 1999
Accepted:
August 04 1999
Citation
Valerie A. Kickhoefer, Amara C. Siva, Nancy L. Kedersha, Elisabeth M. Inman, Cristina Ruland, Michel Streuli, Leonard H. Rome; The 193-Kd Vault Protein, Vparp, Is a Novel Poly(Adp-Ribose) Polymerase. J Cell Biol 6 September 1999; 146 (5): 917–928. doi: https://doi.org/10.1083/jcb.146.5.917
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