Home LiteratureArticle Details
PMID: 11239003 Published · ppublish English Comparative Study Journal Article Research Support, U.S. Gov't, Non-P.H.S. Research Support, U.S. Gov't, P.H.S.

Mixed spermatogenic germ cell nuclear extracts exhibit high base excision repair activity.

Nucleic acids research ·Vol. 29 ·No. 6 ·2001-03-15 ·Pages 1366-72

Intano GW, McMahan CA, Walter RB, McCarrey JR, Walter CA

Abstract

Spermatogenic cells exhibit a lower spontaneous mutation frequency than somatic tissues in a lacI transgene and many base excision repair (BER) genes display the highest observed level of expression in the testis. In this study, uracil-DNA glycosylase-initiated BER activity was measured in nuclear extracts prepared from tissues obtained from each of three mouse strains. Extracts from mixed spermatogenic germ cells displayed the greatest activity followed by liver then brain for all three strains, and the activity for a given tissue was consistent among the three strains. Levels of various BER proteins were examined by western blot analyses and found to be consistent with activity levels. Nuclear extracts prepared from purified Sertoli cells, a somatic component of the seminiferous epithelium, exhibited significantly lower activity than mixed spermatogenic cell-type nuclear extracts, thereby suggesting that the high BER activity observed in mixed germ cell nuclear extracts was not a characteristic of all testicular cell types. Nuclear extracts from thymocytes and small intestines were assayed to assess activity in a mitotically active cell type and tissue. Overall, the order of tissues/cells exhibiting the greatest to lowest activity was mixed germ cells > Sertoli cells > thymocytes > small intestine > liver > brain.

MeSH Terms
Animals Blotting, Western Brain Chemistry Cell Extracts/chemistry,pharmacology Cell Nucleus/chemistry DNA/drug effects,genetics,metabolism DNA Ligases/metabolism DNA Polymerase beta/metabolism DNA Repair Liver/chemistry Male Mice Mice, Inbred C57BL Mice, Inbred Strains Nucleotides/metabolism Oligonucleotides/genetics,metabolism Proteins/metabolism Spermatozoa/chemistry,cytology
Chemicals
Cell Extracts Nucleotides Oligonucleotides Proteins DNA DNA Polymerase beta DNA Ligases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Intano G W
Department of Cellular and Structural Biology and Department of Pathology, The University of Texas Health Science Center at San Antonio, San Antonio, TX 78284, USA. [email protected]
McMahan C A
Walter R B
McCarrey J R
Walter C A
References (62)
62 references, click to expand
  1. Accumulation of premutagenic DNA lesions in mice defective in removal of oxidative base damage.
    Proc Natl Acad Sci U S A. 1999 Nov 9;96(23):13300-5 PMID: 10557315
  2. Recognition and processing of damaged DNA.
    J Cell Sci Suppl. 1995;19:73-7 PMID: 8655650
  3. Mmh/Ogg1 gene inactivation results in accumulation of 8-hydroxyguanine in mice.
    Proc Natl Acad Sci U S A. 2000 Apr 11;97(8):4156-61 PMID: 10725358
  4. Uracil-DNA glycosylase (UNG)-deficient mice reveal a primary role of the enzyme during DNA replication.
    Mol Cell. 2000 Jun;5(6):1059-65 PMID: 10912000
  5. Mutation of a BRCT domain selectively disrupts DNA single-strand break repair in noncycling Chinese hamster ovary cells.
    Proc Natl Acad Sci U S A. 2000 Dec 5;97(25):13649-54 PMID: 11095742
  6. Separation of mouse spermatogenic cells by sedimentation velocity. A morphological characterization.
    Dev Biol. 1976 Mar;49(1):119-31 PMID: 176074
  7. Spermatogenic cells of the prepuberal mouse. Isolation and morphological characterization.
    J Cell Biol. 1977 Jul;74(1):68-85 PMID: 874003
  8. Proliferation of Sertoli cells in fetal and postnatal rats: a quantitative autoradiographic study.
    Anat Rec. 1982 Aug;203(4):485-92 PMID: 7137603
  9. DNA ligase activities during hepatocarcinogenesis induced by N-2-acetylaminofluorene.
    Carcinogenesis. 1985 Sep;6(9):1275-7 PMID: 4028324
  10. Cloning of firefly luciferase cDNA and the expression of active luciferase in Escherichia coli.
    Proc Natl Acad Sci U S A. 1985 Dec;82(23):7870-3 PMID: 3906652
  11. Firefly luciferase gene: structure and expression in mammalian cells.
    Mol Cell Biol. 1987 Feb;7(2):725-37 PMID: 3821727
  12. Tissue-specific variation in repair activity for 3-methyladenine in DNA in two stocks of mice.
    Mutat Res. 1988 Mar-Apr;207(3-4):165-9 PMID: 2451783
  13. Difference in the expression level of DNA polymerase beta among mouse tissues: high expression in the pachytene spermatocyte.
    Exp Cell Res. 1989 Mar;181(1):169-80 PMID: 2917600
  14. DNA ligases from rat liver. Purification and partial characterization of two molecular forms.
    Biochemistry. 1990 Jun 26;29(25):6009-17 PMID: 2383569
  15. Molecular cloning of the human XRCC1 gene, which corrects defective DNA strand break repair and sister chromatid exchange.
    Mol Cell Biol. 1990 Dec;10(12):6160-71 PMID: 2247054
  16. DNA damage metabolism and aging.
    Mutat Res. 1990 Sep-Nov;237(5-6):189-210 PMID: 2079959
  17. Mammalian beta-polymerase promoter: large-scale purification and properties of ATF/CREB palindrome binding protein from bovine testes.
    Biochemistry. 1991 Jun 25;30(25):6296-305 PMID: 1829381
  18. Spectra of spontaneous and mutagen-induced mutations in the lacI gene in transgenic mice.
    Proc Natl Acad Sci U S A. 1991 Sep 15;88(18):7958-62 PMID: 1832771
  19. Differential expression of poly(ADP-ribose) polymerase and DNA polymerase beta in rat tissues.
    Exp Cell Res. 1991 Nov;197(1):66-74 PMID: 1915664
  20. Cloning and expression of APE, the cDNA encoding the major human apurinic endonuclease: definition of a family of DNA repair enzymes.
    Proc Natl Acad Sci U S A. 1991 Dec 15;88(24):11450-4 PMID: 1722334
  21. DNA polymerase-beta and poly(ADP)ribose polymerase mRNAs are differentially expressed during the development of male germinal cells.
    Biol Reprod. 1992 Feb;46(2):201-7 PMID: 1536896
  22. Cloning and characterization of a mouse 3-methyladenine/7-methyl-guanine/3-methylguanine DNA glycosylase cDNA whose gene maps to chromosome 11.
    Carcinogenesis. 1993 Feb;14(2):175-81 PMID: 8435858
  23. DNA repair helicase: a component of BTF2 (TFIIH) basic transcription factor.
    Science. 1993 Apr 2;260(5104):58-63 PMID: 8465201
  24. Purification, culture, and fractionation of spermatogenic cells.
    Methods Enzymol. 1993;225:84-113 PMID: 8231890
  25. Dual role of TFIIH in DNA excision repair and in transcription by RNA polymerase II.
    Nature. 1994 Apr 21;368(6473):769-72 PMID: 8152490
  26. The use of shuttle vectors for mutation analysis in transgenic mice and rats.
    Mutat Res. 1994 Jun 1;307(2):461-78 PMID: 7514720
  27. Deletion of a DNA polymerase beta gene segment in T cells using cell type-specific gene targeting.
    Science. 1994 Jul 1;265(5168):103-6 PMID: 8016642
  28. Proliferating cell nuclear antigen-dependent abasic site repair in Xenopus laevis oocytes: an alternative pathway of base excision DNA repair.
    Mol Cell Biol. 1994 Sep;14(9):6187-97 PMID: 7915006
  29. Characterization of the XRCC1-DNA ligase III complex in vitro and its absence from mutant hamster cells.
    Nucleic Acids Res. 1995 Dec 11;23(23):4836-43 PMID: 8532526
  30. Requirement of mammalian DNA polymerase-beta in base-excision repair.
    Nature. 1996 Jan 11;379(6561):183-6 PMID: 8538772
  31. Repair-deficient 3-methyladenine DNA glycosylase homozygous mutant mouse cells have increased sensitivity to alkylation-induced chromosome damage and cell killing.
    EMBO J. 1996 Feb 15;15(4):945-52 PMID: 8631315
  32. Differential expression of the apurinic / apyrimidinic endonuclease (APE/ref-1) multifunctional DNA base excision repair gene during fetal development and in adult rat brain and testis.
    Mutat Res. 1996 Apr 2;362(3):237-48 PMID: 8637502
  33. Two pathways for base excision repair in mammalian cells.
    J Biol Chem. 1996 Apr 19;271(16):9573-8 PMID: 8621631
  34. DNA ligase I is required for fetal liver erythropoiesis but is not essential for mammalian cell viability.
    Nat Genet. 1996 Aug;13(4):489-91 PMID: 8696349
  35. The redox/DNA repair protein, Ref-1, is essential for early embryonic development in mice.
    Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8919-23 PMID: 8799128
  36. Xrcc-1 expression during male meiosis in the mouse.
    Biol Reprod. 1996 Sep;55(3):630-5 PMID: 8862781
  37. XRCC1 polypeptide interacts with DNA polymerase beta and possibly poly (ADP-ribose) polymerase, and DNA ligase III is a novel molecular 'nick-sensor' in vitro.
    Nucleic Acids Res. 1996 Nov 15;24(22):4387-94 PMID: 8948628
  38. Reconstitution of DNA base excision-repair with purified human proteins: interaction between DNA polymerase beta and the XRCC1 protein.
    EMBO J. 1996 Dec 2;15(23):6662-70 PMID: 8978692
  39. An alternative splicing event which occurs in mouse pachytene spermatocytes generates a form of DNA ligase III with distinct biochemical properties that may function in meiotic recombination.
    Mol Cell Biol. 1997 Feb;17(2):989-98 PMID: 9001252
  40. Lack of phenotypic alteration of hmUra-DNA glycosylase-deficient hamster cells exposed to DNA-damaging agents.
    Mutat Res. 1997 Mar 21;374(2):287-95 PMID: 9100852
  41. XRCC1 protein interacts with one of two distinct forms of DNA ligase III.
    Biochemistry. 1997 Apr 29;36(17):5207-11 PMID: 9136882
  42. Second pathway for completion of human DNA base excision-repair: reconstitution with purified proteins and requirement for DNase IV (FEN1).
    EMBO J. 1997 Jun 2;16(11):3341-8 PMID: 9214649
  43. Base transitions dominate the mutational spectrum of a transgenic reporter gene in MSH2 deficient mice.
    Oncogene. 1997 Jul 10;15(2):123-9 PMID: 9244348
  44. Involvement of XRCC1 and DNA ligase III gene products in DNA base excision repair.
    J Biol Chem. 1997 Sep 19;272(38):23970-5 PMID: 9295348
  45. Base excision repair deficient mice lacking the Aag alkyladenine DNA glycosylase.
    Proc Natl Acad Sci U S A. 1997 Nov 25;94(24):13087-92 PMID: 9371804
  46. XRCC1 is specifically associated with poly(ADP-ribose) polymerase and negatively regulates its activity following DNA damage.
    Mol Cell Biol. 1998 Jun;18(6):3563-71 PMID: 9584196
  47. DNA polymerase beta conducts the gap-filling step in uracil-initiated base excision repair in a bovine testis nuclear extract.
    J Biol Chem. 1995 Jan 13;270(2):949-57 PMID: 7822335
  48. Testis and somatic Xrcc-1 DNA repair gene expression.
    Somat Cell Mol Genet. 1994 Nov;20(6):451-61 PMID: 7892645
  49. Reconstitution of the DNA base excision-repair pathway.
    Curr Biol. 1994 Dec 1;4(12):1069-76 PMID: 7535646
  50. Purification and characterization of DNA ligase III from bovine testes. Homology with DNA ligase II and vaccinia DNA ligase.
    J Biol Chem. 1995 Apr 21;270(16):9683-90 PMID: 7721901
  51. Mammalian base excision repair and DNA polymerase beta.
    Mutat Res. 1998 Jun;407(3):203-15 PMID: 9653447
  52. The influence of DNA glycosylases on spontaneous mutation.
    Mutat Res. 1998 May 25;400(1-2):33-44 PMID: 9685578
  53. Mutation frequency declines during spermatogenesis in young mice but increases in old mice.
    Proc Natl Acad Sci U S A. 1998 Aug 18;95(17):10015-9 PMID: 9707592
  54. Alkylpurine-DNA-N-glycosylase knockout mice show increased susceptibility to induction of mutations by methyl methanesulfonate.
    Mol Cell Biol. 1998 Oct;18(10):5828-37 PMID: 9742100
  55. A murine AP-endonuclease gene-targeted deficiency with post-implantation embryonic progression and ionizing radiation sensitivity.
    Mutat Res. 1998 Oct 21;409(1):17-29 PMID: 9806499
  56. Base excision repair of oxidative DNA damage activated by XPG protein.
    Mol Cell. 1999 Jan;3(1):33-42 PMID: 10024877
  57. Requirement for the Xrcc1 DNA base excision repair gene during early mouse development.
    Dev Biol. 1999 Apr 15;208(2):513-29 PMID: 10191063
  58. Role of DNA polymerase beta in the excision step of long patch mammalian base excision repair.
    J Biol Chem. 1999 May 14;274(20):13741-3 PMID: 10318775
  59. Solution structure of the single-strand break repair protein XRCC1 N-terminal domain.
    Nat Struct Biol. 1999 Sep;6(9):884-93 PMID: 10467102
  60. Mammalian DNA ligase III: molecular cloning, chromosomal localization, and expression in spermatocytes undergoing meiotic recombination.
    Mol Cell Biol. 1995 Oct;15(10):5412-22 PMID: 7565692
  61. Expression of the DNA repair gene XRCC1 in baboon tissues.
    Mutat Res. 1995 Nov;348(3):111-6 PMID: 8524362
  62. A cell cycle-specific requirement for the XRCC1 BRCT II domain during mammalian DNA strand break repair.
    Mol Cell Biol. 2000 Jan;20(2):735-40 PMID: 10611252
Article Info
Journal
Nucleic acids research
Abbr.
Nucleic Acids Res
ISSN
1362-4962
DOI
10.1093/nar/29.6.1366
Email
[email protected]
Published
2001-03-15
Pages
1366-72
Language
English
Region
England
NLM ID
0411011
PMCID
PMC29752
Subset
IM
Grants
NIEHS NIH HHS · ES09136 · United States
NIA NIH HHS · AG13560 · United States
NIA NIH HHS · T32 AG000205 · United States
NIA NIH HHS · AG00205 · United States
NIA NIH HHS · AG14674 · United States
External Links
PubMed source DOI: 10.1093/nar/29.6.1366 PMC Full Text View journal details
Analysis Services
Analysis Services

Contact

No. 2 Wenbo Road, Zhangqiu District, Jinan, Shandong

Qilu Normal University · Genelibs Bioinformatics Lab

750 Shunhua Rd, Jinan

2F, Bldg F, University Science Park

Tel: 0531-88819269

WeChat Official Account

Follow our WeChat subscription account for real-time updates and the latest in medical and biological research.


Business Email

E-mail: [email protected]

Copyright © 2026.Genelibs Jinan Honghui Biotechnology Co., Ltd. 鲁ICP备20007159号-2