Abstract
The evolutionary conservation of DNA polymerase alpha was assessed by immunological and molecular genetic approaches. Four anti-human KB cell DNA polymerase alpha monoclonal antibodies were tested for their ability to recognize a phylogenetically broad array of eukaryotic DNA polymerases. While the single non-neutralizing antibody used in this study recognizes higher mammalian (human, simian, canine, and bovine) polymerases only, three neutralizing antibodies exhibit greater, but variable, extents of cross-reactivity among vertebrate species. The most highly cross-reactive antibody recognizes a unique epitope on a 165-180 kDa catalytic polypeptide in cell lysates from several eukaryotic sources, as distant from man as the amphibians. Genomic Southern hybridization studies with the cDNA of the human DNA polymerase alpha catalytic polypeptide identify the existence of many consensus DNA sequences within the DNA polymerase genes of vertebrate, invertebrate, plant and unicellular organisms. These findings illustrate the differential evolutionary conservation of four unique epitopes on DNA polymerase alpha among vertebrates and the conservation of specific genetic sequences, presumably reflective of critical functional domains, in the DNA polymerase genes from a broad diversity of living forms.
MeSH Terms
Animals
Antibodies, Monoclonal
Base Sequence
Catalysis
Cattle
Cell Line
Cloning, Molecular
DNA Polymerase II/genetics,immunology,isolation & purification
Dogs
Ducks
Epitopes/genetics,isolation & purification
Eukaryotic Cells
Fishes
Genes
Humans
Lizards
Mice
Nucleic Acid Conformation
Papio
Phylogeny
Xenopus laevis
Chemicals
Antibodies, Monoclonal
Epitopes
DNA Polymerase II
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Miller M A
Department of Pathology, Stanford University School of Medicine, CA 94305.
Korn D
Wang T S
References (22)
22 references, click to expand
-
Immunological identification of two adenovirus 2-induced early proteins possibly involved in cell transformation.
Nature. 1976 Nov 18;264(5583):263-6
PMID: 1004548
-
DNA polymerase-alpha. Purification and structural characterization of the near homogeneous enzyme from human KB cells.
J Biol Chem. 1977 Sep 25;252(18):6528-35
PMID: 893425
-
Eukaryotic DNA polymerase alpha. Structural analysis of the enzyme from regenerating rat liver.
J Biol Chem. 1980 Mar 10;255(5):2114-22
PMID: 7354082
-
Chick embryo DNA polymerase alpha. Polypeptide components and their microheterogeneity.
J Biol Chem. 1982 Apr 25;257(8):4484-9
PMID: 7068647
-
10 S DNA polymerase alpha of calf thymus shows a microheterogeneity in its large polypeptide component.
J Biol Chem. 1982 Jun 25;257(12):7172-7
PMID: 7085621
-
Preparation and preliminary characterization of monoclonal antibodies against human DNA polymerase alpha.
J Biol Chem. 1982 Jul 25;257(14):8386-90
PMID: 7085672
-
Isolation of an intact DNA polymerase-primase from embryos of Drosophila melanogaster.
Proc Natl Acad Sci U S A. 1983 Apr;80(8):2221-5
PMID: 6403945
-
Association of DNA primase with the beta/gamma subunits of DNA polymerase alpha from Drosophila melanogaster embryos.
J Biol Chem. 1983 Aug 10;258(15):9037-9
PMID: 6409898
-
Mutations in the herpes simplex virus DNA polymerase gene conferring hypersensitivity to aphidicolin.
Nucleic Acids Res. 1983 Aug 11;11(15):5287-97
PMID: 6308578
-
A DNA primase from mouse cells. Purification and partial characterization.
J Biol Chem. 1983 Aug 25;258(16):9845-9
PMID: 6885774
-
A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity.
Anal Biochem. 1983 Jul 1;132(1):6-13
PMID: 6312838
-
DNA primase from KB cells. Characterization of a primase activity tightly associated with immunoaffinity-purified DNA polymerase-alpha.
J Biol Chem. 1984 Feb 10;259(3):1854-65
PMID: 6693436
-
Immunoaffinity purification and properties of a high molecular weight calf thymus DNA alpha-polymerase.
Biochemistry. 1984 Apr 24;23(9):1895-9
PMID: 6722130
-
Purification of a DNA polymerase-DNA primase complex from calf thymus glands.
J Biol Chem. 1984 Dec 10;259(23):14679-87
PMID: 6094569
-
Polypeptide structure of DNA primase from a yeast DNA polymerase-primase complex.
J Biol Chem. 1985 Jun 10;260(11):7102-7
PMID: 3888995
-
Assignment of the gene for human DNA polymerase alpha to the X chromosome.
Proc Natl Acad Sci U S A. 1985 Aug;82(16):5270-4
PMID: 2410918
-
DNA sequence of the region in the genome of herpes simplex virus type 1 containing the genes for DNA polymerase and the major DNA binding protein.
Nucleic Acids Res. 1985 Nov 25;13(22):8143-63
PMID: 2999714
-
Isolation of the gene encoding yeast DNA polymerase I.
Cell. 1985 Nov;43(1):369-77
PMID: 3907855
-
Role of DNA polymerase alpha and DNA primase in simian virus 40 DNA replication in vitro.
Proc Natl Acad Sci U S A. 1986 May;83(9):2869-73
PMID: 3010320
-
A single-base change within the DNA polymerase locus of herpes simplex virus type 2 can confer resistance to aphidicolin.
J Virol. 1987 Feb;61(2):388-94
PMID: 3027369
-
Related functional domains in virus DNA polymerases.
EMBO J. 1987 Jan;6(1):169-75
PMID: 3034575
-
Human DNA polymerase alpha gene expression is cell proliferation dependent and its primary structure is similar to both prokaryotic and eukaryotic replicative DNA polymerases.
EMBO J. 1988 Jan;7(1):37-47
PMID: 3359994