Studies on a mammalian cell protein (P8) with affinity for DNA in vitro

Tsai, R. L.; Green, Howard

doi:10.1016/0022-2836(73)90344-6

Cited by 91 publications

(28 citation statements)

References 16 publications

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“…Proteins of this type have been isolated first from T4 phage, the gene 32 protein (Alberts & Frey, 1970) and then also from other organisms including E. ¢oli (Abdel-Monem & HoffmannBerling, 1976;Molineux & Gefter, 1974), calf thymus (Herrick & Alberts, 1975 a, b), human fibroblasts (Tsai & Green, 1973), mouse ascites (Otto et al, 1977) and others. These proteins are present in cells in great abundance suggesting a structural rather than an enzymic role.…”

Section: Discussionmentioning

confidence: 99%

“…Indeed, a previous report has described a nucleic acid-binding protein from avian sarcoma virus-lransformed cells with unwinding features which appeared to improve in vitro DNA synthesis (Hung & Lee, 1976, 1977. Nucleic acid-unwinding proteins have been 0022-1317/81/0000-4503 $02.00 ~ 1981 SGM reported in many bacterial and mammalian systems, and appear to be important for DNA replication and genetic recombination (Fleissner & Tress, 1973;Herrick & Alberts, 1975 a, b;Otto et al, 1977;Tsai & Green, 1973;von Hippel & McGhee, 1972). Binding proteins which facilitate RNA conformational changes have not been described extensively.…”

Section: Introductionmentioning

confidence: 99%

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Properties of the Avian Viral Protein p12

Sykora

Moelling

1981

Journal of General Virology

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SUMMARYThe avian RNA tumour virus structural protein p12 was purified from avian myeloblastosis virus (AMV) by nucleic acid affinity chromatography to apparent homogeneity as judged from SDS-polyacrylamide gel electrophoresis. A filterbinding assay was used for the identification of pl2. High concentrations of pl2 precipitated nucleic acids out of solution in the absence of MgC12. Binding of p12 to single-stranded nucleic acids protected them from digestion with nucleases and resulted in a hyperchromic effect. These phenomena were reversible in the presence of salt. The affinity of p12 to nucleic acids was determined by competing for the binding of pl2 to denatured radioactive DNA by various other nucleic acids. It was found that p 12 bound preferentially to single-stranded nucleic acids and showed a higher affinity to poly(rI) than to poly(rC) and poly(rA). Purified RNA-dependent DNA polymerase activity from AMV was stimulated up to sixfold by p12, depending on the template. Solubilization of RNA in RNA-DNA hybrids by RNase H was inhibited in the presence of p12.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Properties of the Avian Viral Protein p12

Sykora

Moelling

1981

Journal of General Virology

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“…Mechali,unlabeled SV40 DNA was from Dr. J. Feunteun. Relaxed, covalently closed SV40 DNA was prepared by 20 min incubation with nicking-closing enzyme in the conditions described by Champoux (7). Single-stranded M13 [ P] DNA (0.032 wg) first mixed with an unlabeled DNA, was incubated with S25 protein in 20 mM Tris-HCl (pH 8.0), 1 nM EDTA, 2 mM 2-mercaptoethanol, 30 mM NaCl and 5% glycerol (Buffer A) in a total volume of 100 wl, at 370C for 30 min.…”

Section: Introductionmentioning

confidence: 99%

Single-strand DNA binding protein from rat liver: interactions with supercoiled DNA

Bonne¹,

Duguet²,

Recondo³

1980

Nucl Acids Res

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As shown by competition experiments, the single-strand DNA binding protein from normal rat liver (S25) interacts preferentially with supercoiled DNA compared to relaxed DNA duplexes. When followed both by sedimentation analysis and by nitrocellulose filter assay, the binding of S25 to SV40 supercoiled DNA (FI) appears to be non-cooperative. Saturation is reached at a protein to DNA weight ratio of about 2. The S25-DNA complexes prefixed with glutaraldehyde appear as beaded structures having an average of 14 to 16 beads per SV40 DNA molecules. Cross-linking of S25 bound to SV40 DNA by dimethyl suberimidate allows to detect oligomeric structures containing a maximum of twenty monomers of S25. When complexes are treated by glutaraldehyde, 10 % of the genome become resistant against micrococcal nuclease. Moreover, S25 affects the DNA helical structure. Superhelical forms are generated by the association of S25 with SV40 DNA, in the presence of nicking-closing enzyme.

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“…Evidence is emerging that GAPDH is also involved in various other cellular functions, such as DNA binding (39,52), tRNA export (45), uracil DNA glycosylase activity (3, 32), and association with polyribosomes (42). Furthermore, an RNA-unwinding property has been identified in GAPDH which is believed to facilitate translation of mRNAs in the polyribosomes (23).…”

mentioning

confidence: 99%

Specific Phosphorylated Forms of Glyceraldehyde 3-Phosphate Dehydrogenase Associate with Human Parainfluenza Virus Type 3 and Inhibit Viral Transcription In Vitro

Choudhary

Banerjee

2000

J Virol

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). To gain further insight into these molecular interactions, we analyzed the virion-associated GAPDH and La protein using two-dimensional gel electrophoresis and immunoblotting. The GAPDH was resolved into two major and one minor molecular species migrating in the pI range of 7.6 to 8.3, while the La protein was resolved into five molecular species in the pI range of 6.8 to 7.5. The GAPDH isoforms present in the virions were also detected in the cytoplasmic fraction of CV-1 cell extract, albeit as minor species. On the other hand, the multiple molecular forms of La protein as seen within the virions were readily detected in the total CV-1 cell extract. Further analysis of virion-associated GAPDH by in vivo labeling with [ 32 P]orthophosphate revealed the presence of multiple phosphorylated species. The phosphorylated species were able to bind specifically to the viral cis-acting 3 genome sense RNA but failed to bind to the leader sense RNA, as determined by gel mobility shift assay. In contrast, the La protein isoforms present within the virions were not phosphorylated and bound to the viral cis-acting RNAs in a phosphorylation-independent manner. The GAPDH isoforms purified from the CV-1 cell cytoplasmic fraction inhibited viral transcription in vitro. Consistent with this, flag-tagged recombinant GAPDH synthesized by using the vaccinia virus expression system also inhibited viral transcription. Together, these data indicate that specific phosphorylated forms of GAPDH associate with HPIV3 and are involved in the regulation of virus gene expression.Human parainfluenza virus type 3 (HPIV3), a paramyxovirus, is second only to respiratory syncytial virus in causing severe respiratory tract infections in children and young adults (6). The single-stranded RNA genome of HPIV3, 15,461 nucleotides (nt) long, is contained within a helical nucleocapsid (15). Three virus-encoded proteins, the nucleocapsid protein N (68 kDa), the phosphoprotein P (90 kDa), and the RNA polymerase L (257 kDa), are associated with the nucleocapsid to form a transcribing ribonucleoprotein (RNP) complex (1, 15). The N protein enwraps the genome RNA, while the L and P proteins together constitute the RNA-dependent RNA polymerase complex that transcribes and replicates the N-bound genome RNA. During transcription, the RNA polymerase synthesizes a short leader RNA, followed by six capped and polyadenylated mRNAs. During replication, on the other hand, a full-length plus-strand copy of the genome RNA is synthesized which, in turn, serves as the template for synthesis of the minus-strand genome RNA (1, 6, 15). Studies from our laboratory, as well as others, indicate that cellular cytoskeletal proteins such as actin and tubulin are involved in the activation of transcription of several paramyxoviruses (4,9,12,22,34,35). Similarly, cellular RNA binding proteins that interact with the viral cis-acting elements are believed to be involved in the regulation of transcription and replication (25,26,28).The minus-sense genomic 3Ј end and its complementary l...

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Studies on a mammalian cell protein (P8) with affinity for DNA in vitro

Cited by 91 publications

References 16 publications

Properties of the Avian Viral Protein p12

Properties of the Avian Viral Protein p12

Single-strand DNA binding protein from rat liver: interactions with supercoiled DNA

Specific Phosphorylated Forms of Glyceraldehyde 3-Phosphate Dehydrogenase Associate with Human Parainfluenza Virus Type 3 and Inhibit Viral Transcription In Vitro

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