Adeno-associated Virus Type 2 Rep78 Inhibition of PKA and PRKX: Fine Mapping and Analysis of Mechanism

Overview

Journal J Virol

Publisher American Society for Microbiology

Date 2002 Jan 5

PMID 11773379

Citations 7

Authors

Michael Schmidt

John A Chiorini

Sandra Afione

Robert Kotin

Affiliations

Soon will be listed here.

Abstract

Hormones and neurotransmitters utilize cyclic AMP (cAMP) as a second messenger in signal transduction pathways to regulate cell growth and division, differentiation, gene expression, and metabolism. Adeno-associated virus type 2 (AAV-2) nonstructural protein Rep78 inhibits members of the cAMP signal transduction pathway, the protein kinases PKA and PRKX. We mapped the kinase binding and inhibition domain of Rep78 for PRKX to amino acids (aa) 526 to 561 and that for PKA to aa 526 to 621. These polypeptides were as potent as full-length Rep78 in kinase inhibition, which suggests that the kinase-inhibitory domain is entirely contained in these Rep peptides. Steady-state kinetic analysis of Rep78-mediated inhibition of PKA and PRKX showed that Rep78 appears to increase the K(m) value of the peptide kinase substrate, while the maximal velocity of the reaction was unaffected. This indicates that Rep78 acts as a competitive inhibitor with respect to the peptide kinase substrate. We detected homology between a cellular pseudosubstrate inhibitor of PKA, the protein kinase inhibitor PKI, and the PRKX and PKA inhibition domains of Rep78. Due to this homology and the competitive inhibition mechanism of Rep78, we propose that Rep78 inhibits PKA and PRKX kinase activity by pseudosubstrate inhibition.

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References

Schmidt M, Afione S, Kotin R . Adeno-associated virus type 2 Rep78 induces apoptosis through caspase activation independently of p53. J Virol. 2000; 74(20):9441-50. PMC: 112373. DOI: 10.1128/jvi.74.20.9441-9450.2000. View

Dubielzig R, King J, Weger S, Kern A, Kleinschmidt J . Adeno-associated virus type 2 protein interactions: formation of pre-encapsidation complexes. J Virol. 1999; 73(11):8989-98. PMC: 112931. DOI: 10.1128/JVI.73.11.8989-8998.1999. View

CASTO B, Armstrong J, ATCHISON R, HAMMON W . Studies on the relationship between adeno-associated virus type 1 (AAV-1) and adenoviruses. II. Inhibition of adenovirus plaques by AAV; its nature and specificity. Virology. 1967; 33(3):452-8. DOI: 10.1016/0042-6822(67)90120-1. View

Laughlin C, Westphal H, Carter B . Spliced adenovirus-associated virus RNA. Proc Natl Acad Sci U S A. 1979; 76(11):5567-71. PMC: 411690. DOI: 10.1073/pnas.76.11.5567. View

Whitehouse S, Feramisco J, Casnellie J, KREBS E, Walsh D . Studies on the kinetic mechanism of the catalytic subunit of the cAMP-dependent protein kinase. J Biol Chem. 1983; 258(6):3693-701. View

Laughlin C, Tratschin J, Coon H, Carter B . Cloning of infectious adeno-associated virus genomes in bacterial plasmids. Gene. 1983; 23(1):65-73. DOI: 10.1016/0378-1119(83)90217-2. View

Kalderon D, Roberts B, Richardson W, Smith A . A short amino acid sequence able to specify nuclear location. Cell. 1984; 39(3 Pt 2):499-509. DOI: 10.1016/0092-8674(84)90457-4. View

Siegl G, Bates R, Berns K, Carter B, Kelly D, Kurstak E . Characteristics and taxonomy of Parvoviridae. Intervirology. 1985; 23(2):61-73. DOI: 10.1159/000149587. View

Scott J, Glaccum M, FISCHER E, KREBS E . Primary-structure requirements for inhibition by the heat-stable inhibitor of the cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1986; 83(6):1613-6. PMC: 323133. DOI: 10.1073/pnas.83.6.1613. View

10.

Glass D, Cheng H, Kemp B, Walsh D . Differential and common recognition of the catalytic sites of the cGMP-dependent and cAMP-dependent protein kinases by inhibitory peptides derived from the heat-stable inhibitor protein. J Biol Chem. 1986; 261(26):12166-71. View

11.

Redemann B, Mendelson E, Carter B . Adeno-associated virus rep protein synthesis during productive infection. J Virol. 1989; 63(2):873-82. PMC: 247761. DOI: 10.1128/JVI.63.2.873-882.1989. View

12.

Sassone-Corsi P, Visvader J, Ferland L, Mellon P, Verma I . Induction of proto-oncogene fos transcription through the adenylate cyclase pathway: characterization of a cAMP-responsive element. Genes Dev. 1988; 2(12A):1529-38. DOI: 10.1101/gad.2.12a.1529. View

13.

Chejanovsky N, Carter B . Replication of a human parvovirus nonsense mutant in mammalian cells containing an inducible amber suppressor. Virology. 1989; 171(1):239-47. DOI: 10.1016/0042-6822(89)90531-x. View

14.

Hermonat P . The adeno-associated virus Rep78 gene inhibits cellular transformation induced by bovine papillomavirus. Virology. 1989; 172(1):253-61. DOI: 10.1016/0042-6822(89)90127-x. View

15.

Im D, Muzyczka N . The AAV origin binding protein Rep68 is an ATP-dependent site-specific endonuclease with DNA helicase activity. Cell. 1990; 61(3):447-57. DOI: 10.1016/0092-8674(90)90526-k. View

16.

Berns K . Parvovirus replication. Microbiol Rev. 1990; 54(3):316-29. PMC: 372780. DOI: 10.1128/mr.54.3.316-329.1990. View

17.

Khleif S, Myers T, Carter B, Trempe J . Inhibition of cellular transformation by the adeno-associated virus rep gene. Virology. 1991; 181(2):738-41. DOI: 10.1016/0042-6822(91)90909-u. View

18.

Van Patten S, Ng D, Thng J, Angelos K, Smith A, Walsh D . Molecular cloning of a rat testis form of the inhibitor protein of cAMP-dependent protein kinase. Proc Natl Acad Sci U S A. 1991; 88(12):5383-7. PMC: 51877. DOI: 10.1073/pnas.88.12.5383. View

19.

Knighton D, Zheng J, Ten Eyck L, Xuong N, Taylor S, Sowadski J . Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. Science. 1991; 253(5018):414-20. DOI: 10.1126/science.1862343. View

20.

Kemp B, Pearson R . Intrasteric regulation of protein kinases and phosphatases. Biochim Biophys Acta. 1991; 1094(1):67-76. DOI: 10.1016/0167-4889(91)90027-u. View