RNA dependent DNA polymerase (reverse transcriptase) from avian myeloblastosis virus

a zinc metalloenzyme

D. S. Auld, H. Kawaguchi, Dennis M Livingston, B. L. Vallee

Research output: Contribution to journalArticle

96 Citations (Scopus)

Abstract

RNA tumor viruses contain a characteristic RNA dependent DNA polymerase (reverse transcriptase) which has been thought to be related to the induction of leukemia by this virus. A disturbance in a zinc dependent enzyme system was first postulated to account for the demonstrated differences in zinc metabolism of normal and leukemic leukocytes. In order to investigate the relationship between zinc and the initiation of leukemia in chickens by avian myeloblastosis virus the metalloenzyme nature of its reverse transcriptase was examined. The present data show that this protein is a zinc metalloenzyme demonstrating the postulated relationship between zinc and a leukemic process. Paucity of purified enzyme generated the design of a novel system of analysis incorporating microwave induced emission spectrometry combined with gel exclusion chromatography. It provides precision, reproducibility, and remarkable limits of detection on μl samples containing 10-12 to 10-14 g atoms of metal, and is thus orders of magnitude more sensitive than other methods. The chromatograohic fraction with highest enzymatic activity contains 1.8 x 10-11 g atoms of zinc per 1.6 μg of protein, corresponding to either 1.8 or 2.0 g atoms of zinc per mole of enzyme for a molecular weight previously determined either as 1.6 or 1.8 x 105. Copper, iron and manganese are absent, i.e., at or below the limits of detection, 10-13 to 10-14 g atoms. Agents known to chelate zinc inhibit the enzyme, while their nonchelating isomers do not. The data underline the participation of zinc on nucleic acid metabolism and bear importantly upon the lesions that accompany leukemia and zinc deficiency (26 references).

Original languageEnglish (US)
Pages (from-to)2091-2095
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume71
Issue number5
DOIs
StatePublished - Jan 1 1974

Fingerprint

Avian Myeloblastosis Virus
RNA-Directed DNA Polymerase
Zinc
Leukemia
Enzymes
Gel Chromatography
Limit of Detection
Virus Activation
Retroviridae
Manganese
Microwaves
Systems Analysis
Nucleic Acids
Copper
Chickens
Spectrum Analysis
Proteins
Leukocytes
Iron
Molecular Weight

Cite this

RNA dependent DNA polymerase (reverse transcriptase) from avian myeloblastosis virus : a zinc metalloenzyme. / Auld, D. S.; Kawaguchi, H.; Livingston, Dennis M; Vallee, B. L.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 71, No. 5, 01.01.1974, p. 2091-2095.

Research output: Contribution to journalArticle

@article{9629cca10f5c425ab4d2d533e06569de,
title = "RNA dependent DNA polymerase (reverse transcriptase) from avian myeloblastosis virus: a zinc metalloenzyme",
abstract = "RNA tumor viruses contain a characteristic RNA dependent DNA polymerase (reverse transcriptase) which has been thought to be related to the induction of leukemia by this virus. A disturbance in a zinc dependent enzyme system was first postulated to account for the demonstrated differences in zinc metabolism of normal and leukemic leukocytes. In order to investigate the relationship between zinc and the initiation of leukemia in chickens by avian myeloblastosis virus the metalloenzyme nature of its reverse transcriptase was examined. The present data show that this protein is a zinc metalloenzyme demonstrating the postulated relationship between zinc and a leukemic process. Paucity of purified enzyme generated the design of a novel system of analysis incorporating microwave induced emission spectrometry combined with gel exclusion chromatography. It provides precision, reproducibility, and remarkable limits of detection on μl samples containing 10-12 to 10-14 g atoms of metal, and is thus orders of magnitude more sensitive than other methods. The chromatograohic fraction with highest enzymatic activity contains 1.8 x 10-11 g atoms of zinc per 1.6 μg of protein, corresponding to either 1.8 or 2.0 g atoms of zinc per mole of enzyme for a molecular weight previously determined either as 1.6 or 1.8 x 105. Copper, iron and manganese are absent, i.e., at or below the limits of detection, 10-13 to 10-14 g atoms. Agents known to chelate zinc inhibit the enzyme, while their nonchelating isomers do not. The data underline the participation of zinc on nucleic acid metabolism and bear importantly upon the lesions that accompany leukemia and zinc deficiency (26 references).",
author = "Auld, {D. S.} and H. Kawaguchi and Livingston, {Dennis M} and Vallee, {B. L.}",
year = "1974",
month = "1",
day = "1",
doi = "10.1073/pnas.71.5.2091",
language = "English (US)",
volume = "71",
pages = "2091--2095",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "5",

}

TY - JOUR

T1 - RNA dependent DNA polymerase (reverse transcriptase) from avian myeloblastosis virus

T2 - a zinc metalloenzyme

AU - Auld, D. S.

AU - Kawaguchi, H.

AU - Livingston, Dennis M

AU - Vallee, B. L.

PY - 1974/1/1

Y1 - 1974/1/1

N2 - RNA tumor viruses contain a characteristic RNA dependent DNA polymerase (reverse transcriptase) which has been thought to be related to the induction of leukemia by this virus. A disturbance in a zinc dependent enzyme system was first postulated to account for the demonstrated differences in zinc metabolism of normal and leukemic leukocytes. In order to investigate the relationship between zinc and the initiation of leukemia in chickens by avian myeloblastosis virus the metalloenzyme nature of its reverse transcriptase was examined. The present data show that this protein is a zinc metalloenzyme demonstrating the postulated relationship between zinc and a leukemic process. Paucity of purified enzyme generated the design of a novel system of analysis incorporating microwave induced emission spectrometry combined with gel exclusion chromatography. It provides precision, reproducibility, and remarkable limits of detection on μl samples containing 10-12 to 10-14 g atoms of metal, and is thus orders of magnitude more sensitive than other methods. The chromatograohic fraction with highest enzymatic activity contains 1.8 x 10-11 g atoms of zinc per 1.6 μg of protein, corresponding to either 1.8 or 2.0 g atoms of zinc per mole of enzyme for a molecular weight previously determined either as 1.6 or 1.8 x 105. Copper, iron and manganese are absent, i.e., at or below the limits of detection, 10-13 to 10-14 g atoms. Agents known to chelate zinc inhibit the enzyme, while their nonchelating isomers do not. The data underline the participation of zinc on nucleic acid metabolism and bear importantly upon the lesions that accompany leukemia and zinc deficiency (26 references).

AB - RNA tumor viruses contain a characteristic RNA dependent DNA polymerase (reverse transcriptase) which has been thought to be related to the induction of leukemia by this virus. A disturbance in a zinc dependent enzyme system was first postulated to account for the demonstrated differences in zinc metabolism of normal and leukemic leukocytes. In order to investigate the relationship between zinc and the initiation of leukemia in chickens by avian myeloblastosis virus the metalloenzyme nature of its reverse transcriptase was examined. The present data show that this protein is a zinc metalloenzyme demonstrating the postulated relationship between zinc and a leukemic process. Paucity of purified enzyme generated the design of a novel system of analysis incorporating microwave induced emission spectrometry combined with gel exclusion chromatography. It provides precision, reproducibility, and remarkable limits of detection on μl samples containing 10-12 to 10-14 g atoms of metal, and is thus orders of magnitude more sensitive than other methods. The chromatograohic fraction with highest enzymatic activity contains 1.8 x 10-11 g atoms of zinc per 1.6 μg of protein, corresponding to either 1.8 or 2.0 g atoms of zinc per mole of enzyme for a molecular weight previously determined either as 1.6 or 1.8 x 105. Copper, iron and manganese are absent, i.e., at or below the limits of detection, 10-13 to 10-14 g atoms. Agents known to chelate zinc inhibit the enzyme, while their nonchelating isomers do not. The data underline the participation of zinc on nucleic acid metabolism and bear importantly upon the lesions that accompany leukemia and zinc deficiency (26 references).

UR - http://www.scopus.com/inward/record.url?scp=0343905521&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0343905521&partnerID=8YFLogxK

U2 - 10.1073/pnas.71.5.2091

DO - 10.1073/pnas.71.5.2091

M3 - Article

VL - 71

SP - 2091

EP - 2095

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 5

ER -