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Adenovirus DNA replication in vitro is stimulated by RNA from uninfected HeLa cells
Author(s) -
van der Vliet P.C.,
van Dam D.,
Kwant M.M.
Publication year - 1984
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(84)80449-4
Subject(s) - microbiology and biotechnology , dna , dna replication , rna , ribonuclease , hela , biology , dna polymerase , eukaryotic dna replication , in vitro , replication factor c , control of chromosome duplication , chemistry , biochemistry , gene
Adenovirus DNA replication was studied in a partially reconstituted system consisting of purified viral proteins (DNA‐binding protein, precursor terminal protein and Ad DNA polymerase) and a nuclear extract from uninfected HeLa cells. Optimal DNA replication required the presence of a heat‐stable, ribonuclease‐sensitive fraction from the cytosol of uninfected cells. This fraction stimulated the initiation about 3‐fold and the replication of origin fragments 5–10‐fold. Sedimentation analysis indicated the presence of a fast‐sedimenting and a slow‐sedimenting component which complemented each other. At least part of the stimulation was caused by low‐molecular‐mass RNA.