scholarly journals Purification, biochemical characterization and serological analysis of cellular deoxyribonucleic acid polymerases and a reverse transcriptase from spleen of a patient with myelofibrotic syndrome

1977 ◽  
Vol 167 (3) ◽  
pp. 513-524 ◽  
Author(s):  
P Chandra ◽  
L K Steel
Keyword(s):  
Reverse Transcriptase ◽  
Dna Polymerase ◽  
Dna Polymerase Beta ◽  
Human Spleen ◽  
Leukaemia Virus ◽  
Polymerase Gamma ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta ◽  
Dna Polymerase Gamma ◽  
Cellular Dna

The present study describes the separation and purification of a reverse transcriptase and cellular DNA polymerases from the human spleen of a patient with myelofibrotic syndrome. The specific requirements with respect to bivalent cations and template-primers for DNA polymerase-alpha, DNA polymerase-beta and DNA polymerase-gamma, as well as for the reverse transcriptase, are reported. Sedimentation-velocity measurements of the purified enzymes gave values of 150000, 40000, 100000 and 70000 daltons for DNA polymerase-alpha DNA polymerase-beta, DNA polymerase-gamma and the reverse transcriptase respectively. Serological studies have shown that the reverse transcriptase from human spleen is not antigenically related to cellular DNA polymerase-alpha, -beta or -gamma, but is antigenically related to reverse transcriptase from simian sarcoma virus and gibbon-ape leukaemia virus.

scholarly journals Evidence for two forms of reverse transcriptase in human placenta of a patient with breast cancer Purification and biochemical characterization of the enzymes

1981 ◽  
Vol 197 (3) ◽  
pp. 553-563 ◽  
Author(s):  
A Vogel ◽  
P Chandra
Keyword(s):  
Breast Cancer ◽  
Reverse Transcriptase ◽  
Dna Polymerase ◽  
Human Placenta ◽  
Biochemical Characterization ◽  
Human Spleen ◽  
Bivalent Cation ◽  
Polymerase Gamma ◽  
Reverse Transcriptases ◽  
Dna Polymerase Gamma

Two DNA polymerases with properties of viral RNA-directed DNA polymerase were found in the placenta of a patient with breast cancer. Both enzyme activities were purified by column-chromatographic procedures or by preparative isoelectric focusing. The most distinguishing feature of the two enzymes is their specificity to transcribe (rA)n . (dT)12 or (rC)n . (dG)18. The two enzymes differ with respect to their elution profiles from the phosphocellulose column, isoelectric point, molecular weight, bivalent-cation requirements and thermal stability. Serological analysis of the (rA)n . (dT)12-activated enzyme showed that this enzyme is immunologically not related to DNA polymerase-gamma, or to any of the reverse transcriptases purified from retroviruses of avian, murine and subprimate origin. However, the activity of this enzyme was neutralized by antibodies to reverse transcriptase purified from human spleen of a patient with myelofibrosis [Chandra & Steel (1977) Biochem. J. 167, 513-524]. Attempts to purify reverse transcriptase of normal human placenta were repeatedly unsuccessful. Once the crude homogenate of normal placenta was freed from endogenous nucleic acids, no (rC)n. (dG)18-dependent activity cold be detected. U

Participation of deoxyribonucleic acid polymerase alpha in amplification of ribosomal deoxyribonucleic acid in Xenopus laevis

1981 ◽  
Vol 1 (8) ◽  
pp. 680-686
Author(s):  
W Zimmermann ◽  
A Weissbach
Keyword(s):  
Xenopus Laevis ◽  
Dna Polymerase ◽  
Ribosomal Dna ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Polymerase Gamma ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta ◽  
Dna Polymerase Gamma ◽  
Ribosomal Ribonucleic Acid

Aphidicolin, a known inhibitor of eucaryotic deoxyribonucleic acid (DNA) polymerase alpha, efficiently inhibited amplification of ribosomal DNA during oogenesis in Xenopus laevis. DNA polymerase alpha, but not DNA polymerase gamma, as isolated from ovaries, was sensitive to aphidicolin. DNA polymerase beta was not detectable in Xenopus ovary extracts. Therefore, DNA polymerase alpha plays a major role in ribosomal ribonucleic acid gene amplification.

scholarly journals Participation of deoxyribonucleic acid polymerase alpha in amplification of ribosomal deoxyribonucleic acid in Xenopus laevis.

1981 ◽  
Vol 1 (8) ◽  
pp. 680-686 ◽  
Author(s):  
W Zimmermann ◽  
A Weissbach
Keyword(s):  
Xenopus Laevis ◽  
Dna Polymerase ◽  
Ribosomal Dna ◽  
Ribonucleic Acid ◽  
Deoxyribonucleic Acid ◽  
Polymerase Gamma ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta ◽  
Dna Polymerase Gamma ◽  
Ribosomal Ribonucleic Acid

Aphidicolin, a known inhibitor of eucaryotic deoxyribonucleic acid (DNA) polymerase alpha, efficiently inhibited amplification of ribosomal DNA during oogenesis in Xenopus laevis. DNA polymerase alpha, but not DNA polymerase gamma, as isolated from ovaries, was sensitive to aphidicolin. DNA polymerase beta was not detectable in Xenopus ovary extracts. Therefore, DNA polymerase alpha plays a major role in ribosomal ribonucleic acid gene amplification.

scholarly journals Involvement of deoxyribonucleic acid polymerase β in nuclear deoxyribonucleic acid synthesis

1978 ◽  
Vol 173 (1) ◽  
pp. 309-314 ◽  
Author(s):  
T R Butt ◽  
W M Wood ◽  
E L McKay ◽  
R L P Adams
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerase ◽  
Deoxyribonucleic Acid ◽  
Nuclear Dna ◽  
Dna Polymerase Beta ◽  
Cell Nuclei ◽  
High Molecular Weight Dna ◽  
Dna Polymerase Alpha ◽  
Polymerase Beta

The effects on DNA synthesis in vitro in mouse L929-cell nuclei of differential extraction of DNA polymerases alpha and beta were studied. Removal of all measurable DNA polymerase alpha and 20% of DNA polymerase beta leads to a 40% fall in the replicative DNA synthesis. Removal of 70% of DNA polymerase beta inhibits replicative synthesis by 80%. In all cases the nuclear DNA synthesis is sensitive to N-ethylmaleimide and aCTP (arabinosylcytosine triphosphate), though less so than DNA polymerase alpha. Addition of deoxyribonuclease I to the nuclear incubation leads to synthesis of high-molecular-weight DNA in a repair reaction. This occurs equally in nuclei from non-growing or S-phase cells. The former nuclei lack DNA polymerase alpha and the reaction reflects the sensitivity of DNA polymerase beta to inhibiton by N-ethylmaleimide and aCTP.

scholarly journals Study of dogfish (Scyliorhinus caniculus) deoxyribonucleic acid polymerase α and β. Extraction, separation, characterization and changes during spermatogenesis

1980 ◽  
Vol 189 (3) ◽  
pp. 635-639 ◽  
Author(s):  
M Philippe ◽  
P Chevaillier
Keyword(s):  
Dna Polymerase ◽  
Sucrose Gradient ◽  
Deoxyribonucleic Acid ◽  
Dna Polymerases ◽  
Polymerase Activity ◽  
Polymerase Gamma ◽  
Polymerase Beta ◽  
Dna Polymerase Gamma ◽  
Highly Sensitive ◽  
Extraction Separation

DNA polymerase activity was extracted from testis cells of the dogfish Scyliorhinus caniculus. On a sucrose gradient, two main peaks could be separated, corresponding to DNA polymerases beta (3.8 S) and alpha (7.5 S). DNA polymerase gamma could also be detected when poly(A) . (dT)12 was used as template. The properties of alpha and beta polymerases of this primitive vertebrate were similar to those generally described, especially in mammals. The beta enzyme was highly sensitive to N-ethylmaleimide, however, and could use poly(dT) . poly(A) as template. Polymerase alpha was present in spermatogonia, spermatocytes and spermatids. Activity was maximal in spermatocytes. DNA polymerase beta was present in all testis cells with similar activities in spermatogonia and spermatocytes. Decreased activities were observed during spermiogenesis. Some activity remained associated with the chromatin fraction of mature sperm cells.

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