Isolation of Proteins Involved in the Replication of Adenoviral DNA In vitro

Lichy, Jack H.; Nagata, Kyosuke; Friefeld, Beth R.; Enomoto, Teruya; Field, Jeffrey; Guggenheimer, R A; Ikeda, Joh‐E; Horwitz, Marshall S.; Hurwitz, Jerard

doi:10.1101/sqb.1983.047.01.084

Cited by 45 publications

(29 citation statements)

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“…7B,C show the levels of DIG-dUTP incorporated into each band. As a positive control antiserum against B23 and adenovirus DBP was used, since both these proteins have been shown to play a role in viral replication Lichy et al, 1983;Okuwaki et al, 2001). For negative controls the anti-UBF serum was pre-incubated with a mixture of UBF1 and UBF2 expressed in baculovirus, the pre-immune antiserum was added at the same concentration, and finally anti-EGFP monoclonal antibody was also tried (from Santa Cruz).…”

Section: Dbp (mentioning

confidence: 99%

Nucleolar protein upstream binding factor is sequestered into adenovirus DNA replication centres during infection without affecting RNA polymerase I location or ablating rRNA synthesis

Lawrence

McStay

Matthews

2006

Journal of Cell Science

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When human adenovirus infects human cells there is disruption of rRNA biogenesis. This report examines the effect of adenovirus infection on the nucleolar protein, upstream binding factor (UBF) which plays a major role in regulating rRNA synthesis. We determined that early after infection, UBF associates with the replication of viral DNA, preferentially associating with the ends of the linear viral genome, and that addition of anti-UBF serum to in vitro replication assays markedly reduced viral DNA replication. Regions of UBF important to these observations are also established. Interestingly, sequestering the majority of UBF from the nucleolus did not lead to the ablation of rRNA synthesis or the sequestration of RNA pol I. In infected cells the bulk of RNA synthesis was RNA pol I associated and distinct from the location of most of the detectable UBF. We propose that UBF plays a role in viral DNA replication, further strengthening the role of nucleolar antigens in the adenovirus life cycle.

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Section: Dbp (mentioning

confidence: 99%

Nucleolar protein upstream binding factor is sequestered into adenovirus DNA replication centres during infection without affecting RNA polymerase I location or ablating rRNA synthesis

Lawrence

McStay

Matthews

2006

Journal of Cell Science

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“…This model has been proved in 429 (16)(17)(18) and in adenovirus (20). Also in M2, the formation of an initiation complex between the terminal protein and dAMP has been shown (18).…”

Section: Location O F Replication Terminimentioning

confidence: 90%

“…Also in M2, the formation of an initiation complex between the terminal protein and dAMP has been shown (18). Therefore, M2 initiates its DNA replication by protein-priming and elongates by displacing parental single-stranded DNA as in the cases of 429(14-18) and adenovirus (19,20). In addition to these Bacillus subtilis phages and adenovirus, formation of a complex between the terminal protein and a deoxyribonucleotide has recently been observed in Escherichia coli and Salmonella typhimurium phage PRD1 (23) and in Streptococcus pneumoniae phage Cp-l (24).…”

Section: Location O F Replication Terminimentioning

confidence: 99%

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Single-strand displacement replication of bacteriophage M2 DNA.

Matsumoto

Hirokawa

1986

J. Gen. Appl. Microbiol.

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Replicative intermediates of M2 DNA, pulse-labeled with [3H]-thymidine, were isolated and the mode of its replication was examined. Compared with mature M2 DNA the replicative intermediates had the following properties: i) higher sedimentation coefficient in sucrose gradient, ii) higher buoyant density in CsC1 equilibrium density gradient, and iii) sensitivity to nuclease S 1. Two types of intermediates were found by electron microscopic observation. One is unit-length duplex molecules with a single-stranded branch of various lengths. The other is unit-length linear molecules consisting of one single-stranded and one doublestranded region of variable proportions. Distribution of the radioactivity in pulse-labeled completed molecules was analyzed with restriction endonucleases. Both ends of the genome had higher radioactivity, showing that the ends are termini of replication. These results indicate that the replication of M2 DNA proceeds from each end by displacing parental single-stranded DNA.

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“…Using a dot blot assay and different incubation conditions, evidence was obtained that the inhibition is due to immunorecognition of a nuclear factor from HeLa cells. Two nuclear proteins are known to be required for adenovirus DNA replication, nuclear factors 1 and 11. DNA replication in the presence of purified nuclear factor I instead of a crude nuclear extract was only slightly inhibited by the antisera.…”

mentioning

confidence: 96%

Inhibition of adenovirus DNA replication in vitro by autoimmune sera

Pruijn¹,

Kusters²,

Meyling³

et al. 1986

Eur J Biochem

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Sera from patients suffering from autoimmune diseases were analyzed for the presence of antibodies that inhibit adenovirus DNA replication in vitro. DNA replication was studied in a reconstituted system containing purified viral proteins (DNA binding protein, DNA polymerase and the precursor to the terminal protein) and a crude nuclear extract from HeLa cells. About half the autoimmune sera analyzed inhibited DNA replication by more than 50% while only 2 out of 31 control sera showed strong inhibition. The inhibition was caused by the IgG fractions of the sera and was most frequently observed with sera from scleroderma patients. Several lines of evidence indicate that the inhibition is not due to anti-DNA antibodies.The mechanism of inhibition of two strongly inhibitory sera was further investigated. The IgG fractions from these sera blocked DNA chain elongation more than 80% but had no effect on the initiation step or the synthesis of the first 26 nucleotides. Using a dot blot assay and different incubation conditions, evidence was obtained that the inhibition is due to immunorecognition of a nuclear factor from HeLa cells. Two nuclear proteins are known to be required for adenovirus DNA replication, nuclear factors 1 and 11. DNA replication in the presence of purified nuclear factor I instead of a crude nuclear extract was only slightly inhibited by the antisera. In agreement with this, immunorecognition of nuclear factor 1 could not be detected using a dot blot assay. Since nuclear factor I1 is not required in our assay system, these results suggest the existence of another nuclear component involved in adenovirus DNA replication which is neutralized by these antibodies.

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Isolation of Proteins Involved in the Replication of Adenoviral DNA In vitro

Cited by 45 publications

References 0 publications

Nucleolar protein upstream binding factor is sequestered into adenovirus DNA replication centres during infection without affecting RNA polymerase I location or ablating rRNA synthesis

Nucleolar protein upstream binding factor is sequestered into adenovirus DNA replication centres during infection without affecting RNA polymerase I location or ablating rRNA synthesis

Single-strand displacement replication of bacteriophage M2 DNA.

Inhibition of adenovirus DNA replication in vitro by autoimmune sera

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