DNA polymerase alpha
Heterotetrameric polymerase which has intrinsic DNA primase activity. Required for initial DNA synthesis of leading strand at origins and for priming of Okazaki fragments on lagging strand. Probably none of the DNA synthesised by this polymerase remains in the completed DNA strands
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Synonyms |
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Molecular weight |
4 subunits 180, 70-85, 55-60,45-50 kDa |
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Biochemical properties |
•RNA synthesis (primase) •DNA synthesis (polymerase) • details of synthesis (1)
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Motifs |
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Protein interactions |
Sc •Mcm10 (2) (3)
Sp •Ctf4/spt16 in an antagonistic way with each other (4)
Mammals •Rb (5) hyperphosphorylated stimulates polymerase activity. Mechanism (6) •Rpa (7) •P53 (8) •Pp2a g1, cyce g1 cyca s mcm2 g1 ( not s) (9) •Various large elongation complexes containing eg rpa, rfc pcna pol e /d etc eg (10) •Smc1 (11) MAMM •mcm10 (12) (13) • AND1 (13) • GINS ( regulates polymerase activity) (14)
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Modifications |
•phosphorylation
SC (15) in meiosis (16) SP (17)
Mammals (9) (18) (19) |
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Structure |
• archael primase (20) |
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Cellular location and expression |
Dm Disperses from nucleus during mitosis (21)
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Other comments |
Mammals •Some effects on telomere length in mouse (22)
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p180 |
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Synonyms |
(Sc) Cdc17, Crt5, Hpr3 |
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Biochemical properties |
•Catalytic polymerase subunit |
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Motifs |
•Nls (3) (23) |
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Interactions |
Sc •Spt16/Pob3 (24) •Pol32 (pol delta subunit) (25)
Mammals •T ag (26) although some confusion on subunits Rb involved in this interaction. Others suggest 70kDa subunit involved (27) . Note that this interaction is species specific. • CaMBP (28) • Parp (29)
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Modifications |
Mammals •cdc2 dependent phosphorylation in mitosis (19) not seen pombe
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Cellular location and expression |
Sc •Protein levels constant with the cell cycle (30) –
Sp •3x variation s/g2 (31) (32)
Mammals •Stays constant levels throughout (33)
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Other comments |
Sc •No cross complementation with pombe enzyme (31) •Mutations affect telomere length (34) • needed in BIR (35)
Dm • e2f sites (36) here has both positive and negative roles (37) and DREF sites (36) • reduction in the absence ATR gives apoptosis/ developmental defects (38)
Mammals •e2f sites mouse (39) • dehydroaltenusin inhibits ( species specific) (40)
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p73 (Sc p85) |
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Biochemical properties |
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Motifs |
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Interactions |
Sp •Orc 2 (41) – suggest this is the loading subunit and 2 modes loading: early through orc or late (bulk) through cdc45
Mammals •Cdc45 (42) (43) • role in RB interaction (6)
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Modifications |
Sc • phosphorylated as orc footprint changes (44) needs 180 for the phosphorylation (15)
Dm •In drosophila phosphorylated on switch to normal cell cycles (after 2h)
Mammals •Mouse Phosphorylated g2/m (45)
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Cellular location and expression |
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Other comments |
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p60 |
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Biochemical properties |
•Primase assist/ transfer between primase and polymerase (ie definition primer length) (46) (42)
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Motifs |
•NLS (47)
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Interactions |
• human primase has required iron sulphur cluster (48) |
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Modifications |
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Cellular location and expression |
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Other comments |
• archael structure (Lao-Sirieix et al., 2005, Nat Struct Mol Biol, 12, 1137-44) |
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p50 |
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Biochemical properties |
•primase (46) |
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Motifs |
•No NLS – relies on 60 for transport?
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Interactions |
archae • rfc (49) |
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Modifications |
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Cellular location and expression |
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Other comments |
Sc •Suggested that some occurs free from complex (50) archae • 1 species has high fidelity primase that synthesizes a mixed RNA/DNA primer (51) |
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Last modified |
14 April 08 |
1. Beckman,J., Kincaid,K., Hocek,M., Spratt,T., Engels,J., Cosstick,R. and Kuchta,R.D. (2007) Human DNA polymerase alpha uses a combination of positive and negative selectivity to polymerize purine dNTPs with high fidelity. Biochemistry 46, 448-460.
2. Fien,K., Cho,Y.S., Lee,J.K., Raychaudhuri,S., Tappin,I. and Hurwitz,J. (2004) Primer utilization by DNA polymerase alpha-primase is influenced by its interaction with Mcm10p. J Biol Chem 279, 16144-16153.
3. Ricke,R.M. and Bielinsky,A.K. (2006) A conserved Hsp10-like domain in Mcm10 is required to stabilize the catalytic subunit of DNA polymerase-alpha in budding yeast. J Biol Chem 281, 18414-18425.
4. Zhou,Y. and Wang,T.S. (2004) A coordinated temporal interplay of nucleosome reorganization factor, sister chromatin cohesion factor, and DNA polymerase alpha facilitates DNA replication. Mol Cell Biol 24, 9568-9579.
5. Takemura,M., Yamamoto,T., Kitagawa,M., Taya,Y., Akiyama,T., Asahara,H., Linn,S., Suzuki,S., Tamai,K. and Yoshida,S. (2001) Stimulation of DNA polymerase alpha activity by cdk2-phosphorylated Rb protein. Biochem Biophys Res Commun 282, 984-990.
6. Takemura,M., Yoshida,S., Akiyama,T., Kitagawa,M. and Yamada,Y. (2006) Role of the second-largest subunit of DNA polymerase alpha in the interaction between the catalytic subunit and hyperphosphorylated retinoblastoma protein in late S phase. Biochim Biophys Acta 1764, 1447-1453.
7. Maga,G., Frouin,I., Spadari,S. and Hubscher,U. (2001) Replication protein A as a "fidelity clamp" for DNA polymerase alpha. J Biol Chem 276, 18235-18242.
8. Melle,C. and Nasheuer,H.P. (2002) Physical and functional interactions of the tumor suppressor protein p53 and DNA polymerase alpha-primase. Nucleic Acids Res 30, 1493-1499.
9. Dehde,S., Rohaly,G., Schub,O., Nasheuer,H.P., Bohn,W., Chemnitz,J., Deppert,W. and Dornreiter,I. (2001) Two immunologically distinct human DNA polymerase alpha-primase subpopulations are involved in cellular DNA replication. Mol Cell Biol 21, 2581-2593.
10. Maga,G. and Hubscher,U. (1996) DNA replication machinery: functional characterization of a complex containing DNA polymerase alpha, DNA polymerase delta, and replication factor C suggests an asymmetric DNA polymerase dimer. Biochemistry 35, 5764-5777.
11. Ryu,M.J., Kim,B.J., Lee,J.W., Lee,M.W., Choi,H.K. and Kim,S.T. (2006) Direct interaction between cohesin complex and DNA replication machinery. Biochem Biophys Res Commun 341, 770-775.
12. Chattopadhyay,S. and Bielinsky,A.K. (2007) Human Mcm10 regulates the catalytic subunit of DNA polymerase-alpha and prevents DNA damage during replication. Mol Biol Cell 18, 4085-4095.
13. Zhu,W., Ukomadu,C., Jha,S., Senga,T., Dhar,S.K., Wohlschlegel,J.A., Nutt,L.K., Kornbluth,S. and Dutta,A. (2007) Mcm10 and And-1/CTF4 recruit DNA polymerase alpha to chromatin for initiation of DNA replication. Genes Dev21, 2288-2299.
14. De Falco,M., Ferrari,E., De Felice,M., Rossi,M., Hubscher,U. and Pisani,F.M. (2007) The human GINS complex binds to and specifically stimulates human DNA polymerase alpha-primase. EMBO Rep 8, 99-103.
15. Ferrari,M., Lucchini,G., Plevani,P. and Foiani,M. (1996) Phosphorylation of the DNA polymerase alpha-primase B subunit is dependent on its association with the p180 polypeptide. J Biol Chem 271, 8661-8666.
16. Foiani,M., Nadjar-Boger,E., Capone,R., Sagee,S., Hashimshoni,T. and Kassir,Y. (1996) A meiosis-specific protein kinase, Ime2, is required for the correct timing of DNA replication and for spore formation in yeast meiosis. Mol Gen Genet 253, 278-288.
17. Bouvier,D., De Recondo,A.M. and Baldacci,G. (1993) In vivo phosphorylation, mitotic behavior, and nuclear binding of the catalytic subunit of DNA polymerase alpha in fission yeast. Exp Cell Res 207, 41-47.
18. Donaldson,R.W. and Gerner,E.W. (1987) Phosphorylation of a high molecular weight DNA polymerase alpha. Proc Natl Acad Sci U S A 84, 759-763.
19. Nasheuer,H.P., Moore,A., Wahl,A.F. and Wang,T.S. (1991) Cell cycle-dependent phosphorylation of human DNA polymerase alpha. J Biol Chem 266, 7893-7903.
20. Lao-Sirieix,S.H., Nookala,R.K., Roversi,P., Bell,S.D. and Pellegrini,L. (2005) Structure of the heterodimeric core primase. Nat Struct Mol Biol 12, 1137-1144.
21. Melov,S., Vaughan,H. and Cotterill,S. (1992) Molecular characterisation of the gene for the 180 kDa subunit of the DNA polymerase-primase of Drosophila melanogaster. J Cell Sci 102, 847-856.
22. Nakamura,M., Nabetani,A., Mizuno,T., Hanaoka,F. and Ishikawa,F. (2005) Alterations of DNA and chromatin structures at telomeres and genetic instability in mouse cells defective in DNA polymerase alpha. Mol Cell Biol 25, 11073-11088.
23. Bouvier,D. and Baldacci,G. (1995) The N-terminus of fission yeast DNA polymerase alpha contains a basic pentapeptide that acts in vivo as a nuclear localization signal. Mol Biol Cell 6, 1697-1705.
24. Wittmeyer,J. and Formosa,T. (1997) The Saccharomyces cerevisiae DNA polymerase alpha catalytic subunit interacts with Cdc68/Spt16 and with Pob3, a protein similar to an HMG1-like protein. Mol Cell Biol 17, 4178-4190.
25. Huang,M.E., Le Douarin,B., Henry,C. and Galibert,F. (1999) The Saccharomyces cerevisiae protein YJR043C (Pol32) interacts with the catalytic subunit of DNA polymerase alpha and is required for cell cycle progression in G2/M. Mol Gen Genet 260, 541-550.
26. Dornreiter,I., Copeland,W.C. and Wang,T.S. (1993) Initiation of simian virus 40 DNA replication requires the interaction of a specific domain of human DNA polymerase alpha with large T antigen. Mol Cell Biol 13, 809-820.
27. Collins,K.L. and Kelly,T.J. (1991) Effects of T antigen and replication protein A on the initiation of DNA synthesis by DNA polymerase alpha-primase. Mol Cell Biol 11, 2108-2115.
28. Cao,Q.P., McGrath,C.A., Baril,E.F., Quesenberry,P.J. and Reddy,G.P. (1995) The 68 kDa calmodulin-binding protein is tightly associated with the multiprotein DNA polymerase alpha-primase complex in HeLa cells. Biochemistry 34, 3878-3883.
29. Dantzer,F., Nasheuer,H.P., Vonesch,J.L., de Murcia,G. and Menissier-de Murcia,J. (1998) Functional association of poly(ADP-ribose) polymerase with DNA polymerase alpha-primase complex: a link between DNA strand break detection and DNA replication. Nucleic Acids Res 26, 1891-1898.
30. Falconi,M.M., Piseri,A., Ferrari,M., Lucchini,G., Plevani,P. and Foiani,M. (1993) De novo synthesis of budding yeast DNA polymerase alpha and POL1 transcription at the G1/S boundary are not required for entrance into S phase. Proc Natl Acad Sci U S A 90, 10519-10523.
31. Park,H., Davis,R. and Wang,T.S. (1995) Studies of Schizosaccharomyces pombe DNA polymerase alpha at different stages of the cell cycle. Nucleic Acids Res23, 4337-4344.
32. Bouvier,D., Pignede,G., Damagnez,V., Tillit,J., de Recondo,A.M. and Baldacci,G. (1992) DNA polymerase alpha in the fission yeast Schizosaccharomyces pombe: identification and tracing of the catalytic subunit during the cell cycle. Exp Cell Res 198, 183-190.
33. Rytkonen,A.K., Vaara,M., Nethanel,T., Kaufmann,G., Sormunen,R., Laara,E., Nasheuer,H.P., Rahmeh,A., Lee,M.Y., Syvaoja,J.E. and Pospiech,H. (2006) Distinctive activities of DNA polymerases during human DNA replication. FEBS J 273, 2984-3001.
34. Adams Martin,A., Dionne,I., Wellinger,R.J. and Holm,C. (2000) The function of DNA polymerase alpha at telomeric G tails is important for telomere homeostasis. Mol Cell Biol 20, 786-796.
35. Lydeard,J.R., Jain,S., Yamaguchi,M. and Haber,J.E. (2007) Break-induced replication and telomerase-independent telomere maintenance require Pol32. Nature448, 820-823.
36. Hirose,F., Yamaguchi,M. and Matsukage,A. (1994) Repression of regulatory factor for Drosophila DNA replication-related gene promoters by zerknullt homeodomain protein. J Biol Chem 269, 2937-2942.
37. Yamaguchi,M., Hayashi,Y., Hirose,F., Nishimoto,Y. and Matsukage,A. (1997) Distinct roles of E2F recognition sites as positive or negative elements in regulation of the DNA polymerase alpha 180 kDa catalytic subunit gene promoter during Drosophila development. Nucleic Acids Res 25, 3847-3854.
38. LaRocque,J.R., Dougherty,D.L., Hussain,S.K. and Sekelsky,J. (2007) Reducing DNA polymerase alpha in the absence of Drosophila ATR leads to P53-dependent apoptosis and developmental defects. Genetics 176, 1441-1451.
39. Nishikawa,N.S., Izumi,M., Uchida,H., Yokoi,M., Miyazawa,H. and Hanaoka,F. (2000) Cloning and characterization of the 5'-upstream sequence governing the cell cycle-dependent transcription of mouse DNA polymerase alpha 68 kDa subunit gene. Nucleic Acids Res 28, 1525-1534.
40. Maeda,N., Kokai,Y., Ohtani,S., Sahara,H., Kuriyama,I., Kamisuki,S., Takahashi,S., Sakaguchi,K., Sugawara,F., Yoshida,H., Sato,N. and Mizushina,Y. (2007) Anti-tumor effects of dehydroaltenusin, a specific inhibitor of mammalian DNA polymerase alpha. Biochem Biophys Res Commun 352, 390-396.
41. Uchiyama,M. and Wang,T.S. (2004) The B-subunit of DNA polymerase alpha-primase associates with the origin recognition complex for initiation of DNA replication. Mol Cell Biol 24, 7419-7434.
42. Arezi,B. and Kuchta,R.D. (2000) Eukaryotic DNA primase. Trends Biochem Sci 25, 572-576.
43. Kukimoto,I., Igaki,H. and Kanda,T. (1999) Human CDC45 protein binds to minichromosome maintenance 7 protein and the p70 subunit of DNA polymerase alpha. Eur J Biochem 265, 936-943.
44. Foiani,M., Liberi,G., Lucchini,G. and Plevani,P. (1995) Cell cycle-dependent phosphorylation and dephosphorylation of the yeast DNA polymerase alpha-primase B subunit. Mol Cell Biol 15, 883-891.
45. Miyazawa,H., Izumi,M., Tada,S., Takada,R., Masutani,M., Ui,M. and Hanaoka,F. (1993) Molecular cloning of the cDNAs for the four subunits of mouse DNA polymerase alpha-primase complex and their gene expression during cell proliferation and the cell cycle. J Biol Chem 268, 8111-8122.
46. Bakkenist,C.J. and Cotterill,S. (1994) The 50-kDa primase subunit of Drosophila melanogaster DNA polymerase alpha. Molecular characterization of the gene and functional analysis of the overexpressed protein. J Biol Chem 269, 26759-26766.
47. Mizuno,T., Okamoto,T., Yokoi,M., Izumi,M., Kobayashi,A., Hachiya,T., Tamai,K., Inoue,T. and Hanaoka,F. (1996) Identification of the nuclear localization signal of mouse DNA primase: nuclear transport of p46 subunit is facilitated by interaction with p54 subunit. J Cell Sci 109, 2627-2636.
48. Weiner,B.E., Huang,H., Dattilo,B.M., Nilges,M.J., Fanning,E. and Chazin,W.J. (2007) An iron-sulfur cluster in the C-terminal domain of the p58 subunit of human DNA primase. J Biol Chem 282, 33444-33451.
49. Wu,K., Lai,X., Guo,X., Hu,J., Xiang,X. and Huang,L. (2007) Interplay between primase and replication factor C in the hyperthermophilic archaeon Sulfolobus solfataricus. Mol Microbiol 63, 826-837.
50. Santocanale,C., Foiani,M., Lucchini,G. and Plevani,P. (1993) The isolated 48,000-dalton subunit of yeast DNA primase is sufficient for RNA primer synthesis. J Biol Chem 268, 1343-1348.
51. Beck,K. and Lipps,G. (2007) Properties of an unusual DNA primase from an archaeal plasmid. Nucleic Acids Res 35, 5635-5645.