Abstract
We have previously described a low-molecular-weight DNA polymerase (52 kDa) from wheat embryo: DNA polymerase CI (pol CI). This enzyme shares some biochemical properties with animal DNA polymerase β (pol β). In this report, we analyse pol CI in wheat embryo germination. Immunodetection and measurement of the enzyme activity show that wheat pol CI remains at a constant level during germination, whereas dramatic changes of the replicative DNA polymerase A and B activities were previously reported. We observe that the level of pol CI in physiological conditions (embryo germination and dividing cell culture) is in agreement with a pol β-type DNA polymerase. By microsequencing of the electroblotted 52 kDa polypeptide, we determined the sequence of a dodecapeptide from the N-terminal region. A comparative analysis of the N-terminal pol CI peptide with some mammalian pol β sequences shows a clear homology with helix 1 of the N-terminal ssDNA domain (residues 15 to 26) of the rat pol β. Thus, the helical structure of this region should be conserved in the wheat peptide. This represents the first evidence of a partial primary structure of a β-type DNA polymerase in plants.
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References
Abbots J, SenGupta DN, Zmudzka B, Widen SG, Notario V, Wilson SH: Expression of human DNA polymerase beta in E. coli and characterization of the recombinant enzyme. Biochemistry 27: 901–909 (1988).
Benedetto JP, Ech-Chaoui R, Plissonneau J, Laquel P, Litvak S, Castroviejo M: Changes of enzymes and factors involved in DNA synthesis during wheat embryo germination. Plant Mol Biol 31: 1217–1225 (1996).
Bradford MM: A rapid and sensitive method for the quantitation of microgram quantities of protein utilising the principe of protein dye binding. Anal Biochem 72: 248–254 (1976).
Bryant JA, Jenns SM, Francis D: DNA polymerase activity and DNA synthesis in roots of pea (Pisum sativum) seedlings. Phytochemistry 20, 13–15 (1981).
Castroviejo M, Gatius M, Litvak S: A low molecular weight DNA polymerase from wheat embryos. Plant Mol Biol 15: 383–387 (1990).
Castroviejo M, Tharaud D, Tarrago-Litvak L, Litvak S: Multiple DNA polymerases from quiescent wheat embryos. Biochem J 181: 183–191 (1979).
Castroviejo M, Tharaud D, Mocquot B, Litvak S: Factors affecting the onset of DNA synthesis during wheat embryo germination. Biochem J 181: 193–199 (1979).
Chang ML, Brown M, Bollum RJ: Induction of DNA polymerase in mouse L cells. J Mol Biol. 74: 1–8 (1983).
Chiu RW, Baril EF: Nuclear DNA polymerases and the HeLa cell cycle. J Biol Chem. 250: 7951–7957 (1975).
Chivers HJ, Bryant JA: Molecular weights of the major DNA polymerases in a higher plant, Pisum sativum L. Biochem Biophys Rcs Com 110: 632–634 (1983).
Dunham VL, Bryant JA: DNA polymerase activities in healthy and cauliflower mosaic virus-infected turnip (Brassica rapa) plants. Ann Bot 57: 81–89 (1986).
Fukasawa H, Yamaguchi M, Chou MH, Matsumoto H, Matsukage A: Characterization of two DNA polymerases from cauliflower inflorescence. J Biochem 87: 1167–1175 (1980).
Johnston FB, Stern H: Mass isolation of viable wheat embryos. Nature 179: 160–161 (1957).
Kim SJ, Lewis MS, Knutson JR, Porter DK, Kumar A, Wilson SH: Characterisation of the tryptophan fluorescence and hydrodynamic properties of rat DNA polymerase beta. J Mol Biol 244: 224–235 (1994).
Kodama H, Komamine A: Cell cycle studies: Induction of synchrony in suspension cultures of Catharanthus roseus cells. Plant Tissue Culture Manual H3: 1–31 (1995).
Kumar A, Widen SG, Williams KR, Kedar P, Karpel RL, Wilson SH: Studies of the domain structure of mammalian DNA polymerase beta. J Biol Chem 265: 2124–2131 (1990).
Laquel P, Litvak S, Castroviejo M: Mammalian PCNA stimulates the processivity of two wheat embryo DNA polymerases. Plant Physiol 102: 107–114 (1993).
Laquel P, Litvak S, Castroviejo M: Wheat DNA primase. Plant Physiol 105: 69–79 (1994).
Liu D, Prasad R, Wilson SH, De Rose EF, Mullen G: Three dimensional solution structure of the N-terminal domain of DNA polymerase beta and the mapping of the ss-DNA interaction interface. Biochemistry 35: 6188–6200 (1996).
Matsumoto Y, Kim Y: Excision of deoxyribose phosphate residues by DNA polymerase beta during DNA repair. Science 269: 699–701 (1995).
Olivieros M, Yanez RJ, Salas ML, Salas J, Vinuela E, Blanco L: Characterisation of an african swine fever virus 20-kDa DNA polymerase involved in DNA repair. J Biol Chem 49: 30899–30099 (1997).
Pelletier H, Sawaya MR, Wolfe W, Wilson SH, Kraut J: Crystal structures of human DNA polymerase beta complexed with DNA: implication for catalytic mechanism, processivity and fidelity. Biochemistry 35: 12742–12761 (1996).
Prasad R, Widen SG, Singhal RK, Watkins J, Prakash L, Wilson SH: Yeast open reading frame YRC14C encodes a DNA beta-polymerase-like enzyme. Nuc Acids Res 21: 5301–5307 (1993).
Robinson N, and Bryant JA: Development of chromatin bound and soluble DNA polymerases activities during germination of Pisum sativum. Planta 127: 259–272 (1975).
Sakaguchi K, Boyd JB: Purification and characterization of DNA polymerase beta from Drosophila. J Biol Chem 260: 10406–10410 (1985).
Sanathkumar M, Ghosh B, SenGupta DN: Isolation of mammalian pol beta type DNA polymerase in shoots tips of germinated seedlings of IR8 rice. Biochem Mol Biol International 39: 117–126 (1996).
Sawaya MR, Pelletier H, Kumar A, Wilson SH, Kraut J: Crystal structure of rat DNA polymerase beta: evidence for a common polymerase mechanism. Science 264: 1930–1935 (1994).
Sedwick WD, Wang TSF, Korn D: Purification and properties of nuclear and cytoplasmic DNApolymerases from human KB cells. J Biol Chem 247: 6026–5033 (1972).
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Luque, A., Benedetto, J. & Castroviejo, M. Wheat DNA polymerase CI: a homologue of rat DNA polymerase β. Plant Mol Biol 38, 647–654 (1998). https://doi.org/10.1023/A:1006052213299
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DOI: https://doi.org/10.1023/A:1006052213299