DNA-directed and RNA-primed DNA synthesis in microsomal and mitochondrial fractions of normal human lymphocytes

Reitz, M. S.; Smith, RG; Roseberry, E.A.; Gallo, Robert C.

doi:10.1016/0006-291x(74)90635-4

Cited by 22 publications

(7 citation statements)

References 11 publications

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“…There was a peak of acid-precipitable radioactivity in the RNA-DNA hybrid region of the gradient, which shifted to the DNA region of the gradient (1 .45 g/ml) by conditions that hydrolyze RNA but not DNA (alkali and RNase) or that disrupt hydrogen but not covalent bonds (heat and alkali) . These results are in contrast to studies with mammalian RNA-primed DNA-instructed DNA polymerase, which catalyzes synthesis of nascent DNA strands covalently linked to RNA primer molecules, which are unaffected by heat treatment (23) . Fig.…”

Section: Requirements Ofcontrasting

confidence: 90%

RNA-instructed DNA polymerase activity in a cytoplasmic particulate fraction in brains from Guamanian patients.

Viola¹,

Frazier²,

White³

et al. 1975

The Journal of Experimental Medicine

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Nervous system tissues from a number of patients with idiopathic neurological disorders were examined for biochemical evidence of RNA tumor virus infection. RNase-sensitive DNA polymerase activity was found in a cytoplasmic particulate fraction from two patients with Guamanian amyotrophic lateral sclerosis (ALS) but not in brains from two normal U.S. individuals. The buoyant density of the enzyme-containing fraction was 1.16-1.18 g/ml and could be converted to a denser region of the gradient (1.24 g/ml) by treatment with the nonionic surfactant, Sterox. The cation and detergent requirements for the endogenous RNase-sensitive DNA polymerase reaction were determined. The early (5 min) endogenous reverse transcriptase product was analyzed by cesium sulfate gradient centrifugation. RNase- and heat-sensitive RNA-DNA hybrids were detected in the product analysis of two ALS, one Parkinsonism-dementia (PD) brain, and two brains from asymptomatic Chamorros but not in brains from normal U.S. individuals and a number of patients with neuro-psychiatric disorders. The DNA product was a 4.5S heteropolymer that hybridized more extensively to RNA extracted from the enzyme-containing pellet from PD brain as compared to a similar fraction from normal U.S. brain. The DNA product appeared to be unrelated to Rausvher or visna virus 70S RNA as determined by RNA-[-3H]DNA hybridization.

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Section: Requirements Ofcontrasting

confidence: 90%

RNA-instructed DNA polymerase activity in a cytoplasmic particulate fraction in brains from Guamanian patients.

Viola¹,

Frazier²,

White³

et al. 1975

The Journal of Experimental Medicine

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“…We needed to develop the assay not only as a very sensitive one but also one that would distinguish RT from all of the then known cellular DNA polymerases (alpha, beta, and gamma). This became a major objective (Bobrow et al, 1972;Robert et al, 1972;Reitz, Jr et al, 1974;Smith et al, 1975;Robert-Guroff and Gallo, 1977).…”

Section: Discovery Of Lymphokines and Early Hints Of Human Retrovirusesmentioning

confidence: 99%

History of the discoveries of the first human retroviruses: HTLV-1 and HTLV-2

Gallo¹

2005

Oncogene

162

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“…An endogenous RNase-sensitive DNA polymerizing activity has also been found in normal human lymphocytes which have been transformed by phytohemagglutinin, but in this case the isolated DNA polymerase is dif ferent from the leukemic enzyme and is not of viral origin since it does not copy 70S viral RNA [17]. The reaction in this case is RNA-primed and DNA-directed [17]. The in vivo functions of either of these DNA polymerases are unknown.…”

mentioning

confidence: 89%

“…The latter is a high-speed post-mitochondrial pellet which has been characterized to the extent that it contains the endogen ous RNase-sensitive DNA-polymerizing activity and that the DNA prod uct hybridizes with RNAs from murine and primate viruses [2,9]. An endogenous RNase-sensitive DNA polymerizing activity has also been found in normal human lymphocytes which have been transformed by phytohemagglutinin, but in this case the isolated DNA polymerase is dif ferent from the leukemic enzyme and is not of viral origin since it does not copy 70S viral RNA [17]. The reaction in this case is RNA-primed and DNA-directed [17].…”

mentioning

confidence: 99%

<i>In vitro </i>DNA Synthesis on Smooth Membranes Observed by Fluorescence

Cavalieri¹,

Sonenberg²,

Cronin-Sheridan³

et al. 1975

Acta Haematol

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Smooth membranes have been isolated from a human diploid line of lymphocytes. These membranes exhibit an endogenous DNA-synthesizing capability which is partially destroyed by prior treatment with RNase. In order to ascertain the role of the membranes in the DNA synthesis we have examined the conformation of the membrane proteins by observing fluorescence changes of the intrinsic probe, tryptophan. We have observed that on addition of the deoxynucleoside-5’-triphosphates, which permits DNA synthesis, there are fluorescence changes due to the tryptophan residue; when DNA synthesis is prevented by omitting some of the precursor triphosphates, fluorescence changes are absent. These effects have been observed with plasma and nuclear membrane fractions; the former may contain a small fraction of the latter. Similar membrane preparations from non-lymphoid cells do not possess the endogenous DNA-synthesizing system, as shown by the lack of incorporation of radioactive precursors or fluorescence changes.

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DNA-directed and RNA-primed DNA synthesis in microsomal and mitochondrial fractions of normal human lymphocytes

Cited by 22 publications

References 11 publications

RNA-instructed DNA polymerase activity in a cytoplasmic particulate fraction in brains from Guamanian patients.

RNA-instructed DNA polymerase activity in a cytoplasmic particulate fraction in brains from Guamanian patients.

History of the discoveries of the first human retroviruses: HTLV-1 and HTLV-2

<i>In vitro </i>DNA Synthesis on Smooth Membranes Observed by Fluorescence

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