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.

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 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 DNA-metabolizing enzymes in normal human lymphoid cells. VI. Induction of DNA polymerases alpha, beta, and gamma following stimulation with phytohemagglutinin

Blood ◽  
1975 ◽  
Vol 46 (4) ◽  
pp. 509-518
Author(s):  
RJ Mayer ◽  
RG Smith ◽  
RC Gallo
Keyword(s):  
Dna Polymerase ◽  
Mammalian Cells ◽  
Dna Polymerases ◽  
Polymerase Activity ◽  
Lymphoid Cells ◽  
Blood Lymphocytes ◽  
Polymerase Beta ◽  
Normal Human ◽  
Alpha Beta ◽  
Cellular Dna

At least three distinct DNA polymerases, named alpha, beta, and gamma, have been isolated from normal mammalian cells. The function of these enzymes in regard to DNA replication and repair remains unclear. Stimulation of blood lymphocytes with the plant mitogen phytohemagglutinin (PHA), is known to increase total DNA polymerase activity. In this study, we measured the change of each of these activities as lymphocytes intered a mitotic cycle. Aliquots of a pool of normal human blood lymphocytes were incubated with PHA for 0, 24, 48, and 72 hr, respectively, and the various DNA polymerase activities quantitated at each point. No significant DNA polymerase activity was detected in unstimulated cells. Low levels of polymerase beta were found at 24 hr. The average DNA content per cell doubled between 24 and 48 hr, and during this interval all three DNA polymerases increased to easily detectable levels. By far the greatest fractional increase in activity of all three polymerases was seen between 48 and 72 hr, after the average doubling of cellular DNA. In summary, these blood lymphocytes lack significant levels of DNA polymerases; stimulation with PHA induces all three of the major DNA polymerase species. In both these respects, these cells differ from other proliferating mammalian cell systems. The possible significance of this difference is discussed.

scholarly journals DNA-metabolizing enzymes in normal human lymphoid cells. VI. Induction of DNA polymerases alpha, beta, and gamma following stimulation with phytohemagglutinin

Blood ◽  
1975 ◽  
Vol 46 (4) ◽  
pp. 509-518 ◽  
Author(s):  
RJ Mayer ◽  
RG Smith ◽  
RC Gallo
Keyword(s):  
Dna Polymerase ◽  
Mammalian Cells ◽  
Dna Polymerases ◽  
Polymerase Activity ◽  
Lymphoid Cells ◽  
Blood Lymphocytes ◽  
Polymerase Beta ◽  
Normal Human ◽  
Alpha Beta ◽  
Cellular Dna

Abstract At least three distinct DNA polymerases, named alpha, beta, and gamma, have been isolated from normal mammalian cells. The function of these enzymes in regard to DNA replication and repair remains unclear. Stimulation of blood lymphocytes with the plant mitogen phytohemagglutinin (PHA), is known to increase total DNA polymerase activity. In this study, we measured the change of each of these activities as lymphocytes intered a mitotic cycle. Aliquots of a pool of normal human blood lymphocytes were incubated with PHA for 0, 24, 48, and 72 hr, respectively, and the various DNA polymerase activities quantitated at each point. No significant DNA polymerase activity was detected in unstimulated cells. Low levels of polymerase beta were found at 24 hr. The average DNA content per cell doubled between 24 and 48 hr, and during this interval all three DNA polymerases increased to easily detectable levels. By far the greatest fractional increase in activity of all three polymerases was seen between 48 and 72 hr, after the average doubling of cellular DNA. In summary, these blood lymphocytes lack significant levels of DNA polymerases; stimulation with PHA induces all three of the major DNA polymerase species. In both these respects, these cells differ from other proliferating mammalian cell systems. The possible significance of this difference is discussed.

scholarly journals The DNA Polymerase Gamma R953C Mutant Is Associated with Antiretroviral Therapy-Induced Mitochondrial Toxicity

2016 ◽  
Vol 60 (9) ◽  
pp. 5608-5611 ◽  
Author(s):  
Min Li ◽  
Andrea C. Mislak ◽  
Yram Foli ◽  
Esinam Agbosu ◽  
Vivek Bose ◽  
...  
Keyword(s):  
Antiretroviral Therapy ◽  
Dna Polymerase ◽  
Infected Patient ◽  
Polymerase Activity ◽  
Mitochondrial Toxicity ◽  
Wild Type ◽  
Polymerase Gamma ◽  
Dna Polymerase Gamma ◽  
Deoxynucleoside Triphosphate ◽  
Dntp Binding

ABSTRACTWe found a heterozygous C2857T mutation (R953C) in polymerase gamma (Pol-γ) in an HIV-infected patient with mitochondrial toxicity. The R953C Pol-γ mutant binding affinity for dCTP is 8-fold less than that of the wild type. The R953C mutant shows a 4-fold decrease in discrimination of analog nucleotides relative to the wild type. R953 is located on the “O-helix” that forms the substrate deoxynucleoside triphosphate (dNTP) binding site; the interactions of R953 with E1056 and Y986 may stabilize the O-helix and affect polymerase activity.

scholarly journals Inventory and Evolution of Mitochondrion-localized Family A DNA Polymerases in Euglenozoa

Pathogens ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 257 ◽  
Author(s):  
Ryo Harada ◽  
Yoshihisa Hirakawa ◽  
Akinori Yabuki ◽  
Yuichiro Kashiyama ◽  
Moe Maruyama ◽  
...  
Keyword(s):  
Dna Polymerase ◽  
Dna Polymerases ◽  
Dna Polymerase I ◽  
Bacterial Dna ◽  
Polymerase Gamma ◽  
Origin And Evolution ◽  
Dna Polymerase Gamma ◽  
Unique Biology ◽  
Organellar Dna ◽  
Polymerase I

The order Trypanosomatida has been well studied due to its pathogenicity and the unique biology of the mitochondrion. In Trypanosoma brucei, four DNA polymerases, namely PolIA, PolIB, PolIC, and PolID, related to bacterial DNA polymerase I (PolI), were shown to be localized in mitochondria experimentally. These mitochondrion-localized DNA polymerases are phylogenetically distinct from other family A DNA polymerases, such as bacterial PolI, DNA polymerase gamma (Polγ) in human and yeasts, “plant and protist organellar DNA polymerase (POP)” in diverse eukaryotes. However, the diversity of mitochondrion-localized DNA polymerases in Euglenozoa other than Trypanosomatida is poorly understood. In this study, we discovered putative mitochondrion-localized DNA polymerases in broad members of three major classes of Euglenozoa—Kinetoplastea, Diplonemea, and Euglenida—to explore the origin and evolution of trypanosomatid PolIA-D. We unveiled distinct inventories of mitochondrion-localized DNA polymerases in the three classes: (1) PolIA is ubiquitous across the three euglenozoan classes, (2) PolIB, C, and D are restricted in kinetoplastids, (3) new types of mitochondrion-localized DNA polymerases were identified in a prokinetoplastid and diplonemids, and (4) evolutionarily distinct types of POP were found in euglenids. We finally propose scenarios to explain the inventories of mitochondrion-localized DNA polymerases in Kinetoplastea, Diplonemea, and Euglenida.

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.

Sign in / Sign up

Export Citation Format

Share Document