Home LiteratureArticle Details
PMID: 7813449 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Tc1 transposase of Caenorhabditis elegans is an endonuclease with a bipartite DNA binding domain.

The EMBO journal ·Vol. 13 ·No. 24 ·1994-12-15 ·Pages 6125-32

Vos JC, Plasterk RH

Abstract

The Tc1 transposon of Caenorhabditis elegans is a member of the Tc1/mariner family of mobile elements. These elements have inverted terminal repeats that flank a single transposase gene. Here we show that Tc1 transposase, Tc1A, has a bipartite DNA binding domain related to the paired domain of mammalian and Drosophila genes. Both the DNA binding domain of Tc1A and the DNA binding site in the inverted repeat of Tc1 can be divided into two subdomains. Methylation interference studies demonstrate adjacent minor and major groove contacts at the inner part of the binding site by the N-terminal 68 amino acids of the DNA binding domain. In addition, Tc1A amino acids 69-142 are essential for major groove contacts at the outer part of the binding site. Recombinant Tc1A is found to be able to introduce a single strand nick at the 5' end of the transposon in vitro. Furthermore, Tc1A can mediate a phosphoryl transfer reaction. A mutation in a DDE motif abolishes both endonucleolytic and phosphoryl transfer activities, suggesting that Tc1A carries a catalytic core common to retroviral integrases and IS transposases.

MeSH Terms
Animals Base Sequence Binding Sites Caenorhabditis elegans/enzymology DNA Transposable Elements DNA-Binding Proteins/genetics,isolation & purification,metabolism Endonucleases/genetics,metabolism Escherichia coli/genetics Methylation Molecular Sequence Data Nucleotidyltransferases/genetics,isolation & purification,metabolism Repetitive Sequences, Nucleic Acid Substrate Specificity Transposases
Chemicals
DNA Transposable Elements DNA-Binding Proteins Nucleotidyltransferases Tc1 transposase Transposases Endonucleases
Authors & Affiliations
2 authors, click to expand affiliations / ORCID
Vos J C
The Netherlands Cancer Institute, Division of Molecular Biology, Amsterdam.
Plasterk R H
References (32)
32 references, click to expand
  1. Sequencing end-labeled DNA with base-specific chemical cleavages.
    Methods Enzymol. 1980;65(1):499-560 PMID: 6246368
  2. The mechanism of transposition of Tc3 in C. elegans.
    Cell. 1994 Oct 21;79(2):293-301 PMID: 7954797
  3. Target sequences for the C. elegans transposable element Tc1.
    Nucleic Acids Res. 1983 Oct 25;11(20):7137-40 PMID: 6314277
  4. Extrachromosomal copies of transposon Tc1 in the nematode Caenorhabditis elegans.
    Proc Natl Acad Sci U S A. 1984 Jul;81(13):4018-22 PMID: 6330730
  5. Conservation of a large protein domain in the segmentation gene paired and in functionally related genes of Drosophila.
    Cell. 1986 Dec 26;47(6):1033-40 PMID: 2877747
  6. Vectors for selective expression of cloned DNAs by T7 RNA polymerase.
    Gene. 1987;56(1):125-35 PMID: 3315856
  7. Efficient oligonucleotide-directed construction of mutations in expression vectors by the gapped duplex DNA method using alternating selectable markers.
    Nucleic Acids Res. 1989 Jun 26;17(12):4441-54 PMID: 2501754
  8. Mutational analysis of IS10's outside end.
    EMBO J. 1989 Jul;8(7):2101-9 PMID: 2551675
  9. Functional similarities between retroviruses and the IS3 family of bacterial insertion sequences?
    Mol Microbiol. 1990 Oct;4(10):1771-7 PMID: 1963920
  10. Retroviral integrase domains: DNA binding and the recognition of LTR sequences.
    Nucleic Acids Res. 1991 Feb 25;19(4):851-60 PMID: 1850126
  11. The origin of footprints of the Tc1 transposon of Caenorhabditis elegans.
    EMBO J. 1991 Jul;10(7):1919-25 PMID: 1646714
  12. Pax: a murine multigene family of paired box-containing genes.
    Genomics. 1991 Oct;11(2):424-34 PMID: 1685142
  13. Reversal of integration and DNA splicing mediated by integrase of human immunodeficiency virus.
    Science. 1992 Feb 7;255(5045):723-6 PMID: 1738845
  14. Residues critical for retroviral integrative recombination in a region that is highly conserved among retroviral/retrotransposon integrases and bacterial insertion sequence transposases.
    Mol Cell Biol. 1992 May;12(5):2331-8 PMID: 1314954
  15. Identification of amino acid residues critical for endonuclease and integration activities of HIV-1 IN protein in vitro.
    Virology. 1992 Jun;188(2):459-68 PMID: 1585629
  16. Detection of Caenorhabditis transposon homologs in diverse organisms.
    New Biol. 1992 Apr;4(4):382-8 PMID: 1320399
  17. Transpositional recombination: mechanistic insights from studies of mu and other elements.
    Annu Rev Biochem. 1992;61:1011-51 PMID: 1323232
  18. Binding of the IS903 transposase to its inverted repeat in vitro.
    EMBO J. 1992 Sep;11(9):3449-55 PMID: 1324175
  19. Identification of conserved amino acid residues critical for human immunodeficiency virus type 1 integrase function in vitro.
    J Virol. 1992 Nov;66(11):6361-9 PMID: 1404595
  20. Mutational analysis of the integrase protein of human immunodeficiency virus type 2.
    Proc Natl Acad Sci U S A. 1992 Oct 15;89(20):9598-602 PMID: 1409671
  21. Identification of the catalytic and DNA-binding region of the human immunodeficiency virus type I integrase protein.
    Nucleic Acids Res. 1993 Mar 25;21(6):1419-25 PMID: 8464733
  22. Domains of the integrase protein of human immunodeficiency virus type 1 responsible for polynucleotidyl transfer and zinc binding.
    Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3428-32 PMID: 8386373
  23. Mobilization of quiet, endogenous Tc3 transposons of Caenorhabditis elegans by forced expression of Tc3 transposase.
    EMBO J. 1993 Jun;12(6):2513-20 PMID: 8389698
  24. Characterization of the Caenorhabditis elegans Tc1 transposase in vivo and in vitro.
    Genes Dev. 1993 Jul;7(7A):1244-53 PMID: 8391505
  25. DNA sequence recognition by Pax proteins: bipartite structure of the paired domain and its binding site.
    Genes Dev. 1993 Oct;7(10):2048-61 PMID: 8406007
  26. Influence of substrate structure on disintegration activity of Moloney murine leukemia virus integrase.
    J Virol. 1993 Dec;67(12):7077-87 PMID: 8230431
  27. A proposed superfamily of transposase genes: transposon-like elements in ciliated protozoa and a common "D35E" motif.
    Proc Natl Acad Sci U S A. 1994 Feb 1;91(3):942-6 PMID: 8302872
  28. Activities of the feline immunodeficiency virus integrase protein produced in Escherichia coli.
    J Virol. 1994 Mar;68(3):1468-74 PMID: 8107210
  29. Target site choice of the related transposable elements Tc1 and Tc3 of Caenorhabditis elegans.
    Nucleic Acids Res. 1994 Feb 11;22(3):262-9 PMID: 8127662
  30. Intermolecular disintegration and intramolecular strand transfer activities of wild-type and mutant HIV-1 integrase.
    Nucleic Acids Res. 1994 Mar 25;22(6):1037-43 PMID: 8152908
  31. Mobile Minos elements from Drosophila hydei encode a two-exon transposase with similarity to the paired DNA-binding domain.
    Proc Natl Acad Sci U S A. 1994 May 24;91(11):4746-50 PMID: 8197129
  32. Sequence of the C. elegans transposable element Tc1.
    Nucleic Acids Res. 1983 Jun 25;11(12):4201-9 PMID: 6306578
Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
Published
1994-12-15
Pages
6125-32
Language
English
Region
England
NLM ID
8208664
PMCID
PMC395592
Subset
IM
External Links
PubMed source 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