Tyrosine kinase-dependent release of an adenovirus preterminal protein complex from the nuclear matrix

Peter C Angeletti, Jeffrey A. Engler

Research output: Contribution to journalArticle

17 Citations (Scopus)

Abstract

Adenovirus (Ad) replicative complexes form at discrete sites on the nuclear matrix (NM) through the interaction of Ad preterminal protein (pTP). The NM is a highly salt-resistant fibrillar network which is known to anchor transcription, mRNA splicing, and DNA replication complexes. Incubation of rATP with NM to which pTP was bound caused the release of pTP as a pTP-NM complex with a size of 220 to 230 kDa; incubation with 5' adenylylimidodiphosphate (rAMP-PNP) showed no significant release, indicating that rATP hydrolysis was required. With NM extracts, it was shown that a pTP- NM complex which was capable of binding Ad origin DNA could be reconstituted in vitro. A number of high-molecular-weight NM proteins ranging in size from 120 in 200 kDa were identified on Far Western blots for their ability in bind pTP. rATP-dependent release of pTP from the NM was inhibited in a dose- dependent fashion by the addition of tyrosine kinase inhibitors, such as quercetin, methyl-2,5-dihydroxycinnamate, or genistein. NM-mediated phosphorylation of a poly(Glu, Tyr) substrate was also significantly abrogated by the addition of these compounds. rATP-dependent release of Ad DNA termini bound to the NM via pTP was also blocked by the addition of these inhibitors. These results indicate that a tyrosine kinase mechanism controls the release of pTP from its binding sites on the NM. These data support the concept that phosphorylation may play a key role in the modulation of pTP binding sites on the NM.

Original languageEnglish (US)
Pages (from-to)3060-3067
Number of pages8
JournalJournal of virology
Volume70
Issue number5
StatePublished - May 3 1996

Fingerprint

nuclear matrix
Nuclear Matrix
Adenoviridae
Protein-Tyrosine Kinases
tyrosine
phosphotransferases (kinases)
Nuclear Matrix-Associated Proteins
proteins
Protein Binding
Proteins
Binding Sites
Phosphorylation
Far-Western Blotting
Adenylyl Imidodiphosphate
adenovirus pTP protein
Genistein
binding sites
DNA
phosphorylation
Quercetin

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Tyrosine kinase-dependent release of an adenovirus preterminal protein complex from the nuclear matrix. / Angeletti, Peter C; Engler, Jeffrey A.

In: Journal of virology, Vol. 70, No. 5, 03.05.1996, p. 3060-3067.

Research output: Contribution to journalArticle

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