Reverse transcriptase from avian myeloblastosis virus

Houts, G E; Miyagi, M.; Ellis, C; Beard, Dorothy; Beard, J. W.

doi:10.1128/jvi.29.2.517-522.1979

Cited by 66 publications

(24 citation statements)

References 21 publications

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“…Avian myeloblastosis virus (AMV) pp32 was prepared as described by Grandgenett et al (11) by chromatography through phosphocellulose and poly(U)-Sepharose 4B (Pharmacia) columns. The starting material for this preparation was a partially purified high-salt (0.8 M potassium phosphate) eluate fraction from a phosphocellulose column, from which the a,B form of reverse transcriptase from 15 g of AMV was purified as described by Houts et al (14) except that 20 mM ,-glycerophosphate was added to all buffers to inhibit protein phosphatases. The final pp32 protein preparation was concentrated by poly(U)-Sepharose chromatography as described below and stored at -70°C in 40% glycerol buffer.…”

Section: Methodsmentioning

confidence: 99%

Properties of avian sarcoma-leukosis virus pp32-related pol-endonucleases produced in Escherichia coli

Terry

Soltis

Katzman

et al. 1988

J Virol

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The gag-pol precursor protein of the avian sarcoma-leukosis virus is processed into three known pol-encoded mature polypeptides; the 95-and 63-kilodalton (kDa) and a subunits, respectively, of reverse transcriptase and the 32-kDa pp32 protein. The pp32 protein possesses DNA endonuclease activity and is produced from the precursor by two proteolytic cleavage events, one of which removes 4.1 kDa of protein from the C terminus. A 36-kDa protein (p36P°') which retains this C-terminal segment is detectable in small quantities in virions. We have constructed Escherichia coli plasmid clones that express the C-terminal domains of pol corresponding to pp32 and p36. These proteins have been purified by column chromatographic methods to near homogeneity. No significant differences could be detected in the enzymatic properties of the bacterially produced p32P"' and p36&"" proteins. Both possess DNA endonuclease activity and, like the pp32 protein isolated from virions, can cleave near the junction of two tandem avian sarcoma-leukosis virus long terminal repeats in double-stranded supercoiled DNA substrates. In the presence of Mg2+, both p32P"' and viral pp32 cleave either strand of DNA 2 nucleotides 5' to the junction.

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Section: Methodsmentioning

confidence: 99%

Properties of avian sarcoma-leukosis virus pp32-related pol-endonucleases produced in Escherichia coli

Terry

Soltis

Katzman

et al. 1988

J Virol

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“…Avian myeloblastosis virus reverse transcriptase, purified as described in (16), was obtained from Dr. J. W. Beard (Life Sciences Inc. Florida. USA).…”

Section: Methodsmentioning

confidence: 99%

“…Alkylation of the complex was performed as described before (15,25). No changes in the alkylation pattern of native tRNATrp were observed when the buffer described in (15) was replaced by the reverse transcriptase stabilizing buffer described above (16).…”

Section: Methodsmentioning

confidence: 99%

Interactions between avian myeloblastosis reverse transcriptase and tRNA^Trp. Mapping of complexed tRNA with chemicals and nucleases

et al. 1984

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The interactions between beef tRNATrp with avian myeloblastosis reverse transcriptase have been studied by statistical chemical modifications of phosphate (ethylnitrosourea) and cytidine (dimethyl sulfate) residues, as well as by digestion of complexed tRNA by Cobra venom nuclease and Neurospora crassa endonuclease. Results with nucleases and chemicals show that reverse transcriptase interacts preferentially with the D arm, the anticodon stem and the T psi stem. All these regions are located in the outside of the L-shaped structure of tRNA. This domain of interaction is different to that reported previously in the complex of beef tRNA with the cognate aminoacyl-tRNA synthetase (M. Garret et al.; Eur. J. Biochem. In press). Avian reverse transcriptase destabilizes the region of tRNA where most of the tertiary interactions maintaining the structure of tRNA are located.

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“…PolyA'*'-RNA from spruce was reverse transcribed by AMV-reverse transcriptase (HOUTS et al 1979) using ohgo (dT),2_i8 as primer (MANIATIS et al 1982). In order to achieve maximum specific activity, ^^P-dGTP (Amersham, 3,000 Gi/mmol; 10 /^Gi/^g polyA+-RNA) was used without additional non-labeled dGTP.…”

Section: Preparation Of Cdna For Hybridization By Reverse Transcriptimentioning

confidence: 99%

Differential Screening in a cDNA‐Library from Spruce for Clones Associated with Forest Decline Reveals Accumulation of Ribulose‐l,5‐Bisphosphate Carboxylase Small Subunit mRNA

Etscheid

Buschmann

Köhler

et al. 1993

Journal of Phytopathology

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In this work we identified mRNA species in spruce (Picea abies L., Karst), which are expressed in altered concentration if the symptoms of forest decline are present. A cDNA library was constructed and ‘damage‐associated’ clones were identified by differential screening. A procedure for polyA+‐RNA preparation from needles of spruce was established which yielded a RNA fraction of sufficient purity for gel electrophoresis, hybridization and most importantly, for reverse transcription and the consecutive steps of molecular cloning. RNA isolated from a symptomless and a damaged tree was combined and a cDNA library in phage λ gt 10 was constructed. Three series of differential screening were carried out. In the first series radioactively labeled cDNA samples were prepared from poly A+‐RNA from a symptomless tree and a damaged tree. Differential hybridization of 10,000 cDNA clones with the two different samples yielded 37 cDNAs with different levels of hybridization signals. In a second series samples for hybridization tests were prepared from a symptomless and from a damaged tree at three different seasons, July, September, and December. These six different samples were used to search for significant hybridization differences in 2,000 clones. Five clones could be identified which showed hybridization differences independent of the annual season. In a third series, we tried to differentiate between genetic variations of individual trees and differences solely induced by the damage. Two mixed samples were prepared, one from an ensemble of 10 symptomless and one of 10 damaged trees. Six cDNA clones could be identified, which exerted clear differences with the mixed probes from the symptomless ensemble and the damage ensemble, respectively. In Northern hybridizations the cDNA clones could be attributed to mRNAs of well defined length. After sequence analysis of the cDNA clones and homology search in a gene data bank two sequences could be identified. One clone codes for the small subunit of ribulose‐l,5‐bisphosphate carboxylase (Rubisco SS) and another for the subunit II of photosystem I. By quantitative Northern

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Reverse transcriptase from avian myeloblastosis virus

Cited by 66 publications

References 21 publications

Properties of avian sarcoma-leukosis virus pp32-related pol-endonucleases produced in Escherichia coli

Properties of avian sarcoma-leukosis virus pp32-related pol-endonucleases produced in Escherichia coli

Interactions between avian myeloblastosis reverse transcriptase and tRNA^Trp. Mapping of complexed tRNA with chemicals and nucleases

Differential Screening in a cDNA‐Library from Spruce for Clones Associated with Forest Decline Reveals Accumulation of Ribulose‐l,5‐Bisphosphate Carboxylase Small Subunit mRNA

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Reverse transcriptase from avian myeloblastosis virus

Cited by 66 publications

References 21 publications

Properties of avian sarcoma-leukosis virus pp32-related pol-endonucleases produced in Escherichia coli

Properties of avian sarcoma-leukosis virus pp32-related pol-endonucleases produced in Escherichia coli

Interactions between avian myeloblastosis reverse transcriptase and tRNATrp. Mapping of complexed tRNA with chemicals and nucleases

Differential Screening in a cDNA‐Library from Spruce for Clones Associated with Forest Decline Reveals Accumulation of Ribulose‐l,5‐Bisphosphate Carboxylase Small Subunit mRNA

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Interactions between avian myeloblastosis reverse transcriptase and tRNA^Trp. Mapping of complexed tRNA with chemicals and nucleases