Skip to main content
NCBI home page
Genetics logoLink to Genetics
. 1999 May;152(1):291–297. doi: 10.1093/genetics/152.1.291

Molecular characterization of mutant alleles of the DNA repair/basal transcription factor haywire/ERCC3 in Drosophila.

L C Mounkes 1, M T Fuller 1
PMCID: PMC1460603  PMID: 10224261

Abstract

The haywire gene of Drosophila encodes a putative helicase essential for transcription and nucleotide excision repair. A haywire allele encoding a dominant acting poison product, lethal alleles, and viable but UV-sensitive alleles isolated as revertants of the dominant acting poison allele were molecularly characterized. Sequence analysis of lethal haywire alleles revealed the importance of the nucleotide-binding domain, suggesting an essential role for ATPase activity. The viable haync2 allele, which encodes a poison product, has a single amino acid change in conserved helicase domain VI. This mutation results in accumulation of a 68-kD polypeptide that is much more abundant than the wild-type haywire protein.

Full Text

The Full Text of this article is available as a PDF (177.3 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Biggin M. D., Gibson T. J., Hong G. F. Buffer gradient gels and 35S label as an aid to rapid DNA sequence determination. Proc Natl Acad Sci U S A. 1983 Jul;80(13):3963–3965. doi: 10.1073/pnas.80.13.3963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Drapkin R., Reardon J. T., Ansari A., Huang J. C., Zawel L., Ahn K., Sancar A., Reinberg D. Dual role of TFIIH in DNA excision repair and in transcription by RNA polymerase II. Nature. 1994 Apr 21;368(6473):769–772. doi: 10.1038/368769a0. [DOI] [PubMed] [Google Scholar]
  3. Feaver W. J., Svejstrup J. Q., Bardwell L., Bardwell A. J., Buratowski S., Gulyas K. D., Donahue T. F., Friedberg E. C., Kornberg R. D. Dual roles of a multiprotein complex from S. cerevisiae in transcription and DNA repair. Cell. 1993 Dec 31;75(7):1379–1387. doi: 10.1016/0092-8674(93)90624-y. [DOI] [PubMed] [Google Scholar]
  4. Fuller M. T., Regan C. L., Green L. L., Robertson B., Deuring R., Hays T. S. Interacting genes identify interacting proteins involved in microtubule function in Drosophila. Cell Motil Cytoskeleton. 1989;14(1):128–135. doi: 10.1002/cm.970140122. [DOI] [PubMed] [Google Scholar]
  5. Gorbalenya A. E., Koonin E. V., Donchenko A. P., Blinov V. M. Two related superfamilies of putative helicases involved in replication, recombination, repair and expression of DNA and RNA genomes. Nucleic Acids Res. 1989 Jun 26;17(12):4713–4730. doi: 10.1093/nar/17.12.4713. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Guzder S. N., Sung P., Bailly V., Prakash L., Prakash S. RAD25 is a DNA helicase required for DNA repair and RNA polymerase II transcription. Nature. 1994 Jun 16;369(6481):578–581. doi: 10.1038/369578a0. [DOI] [PubMed] [Google Scholar]
  7. Hays T. S., Deuring R., Robertson B., Prout M., Fuller M. T. Interacting proteins identified by genetic interactions: a missense mutation in alpha-tubulin fails to complement alleles of the testis-specific beta-tubulin gene of Drosophila melanogaster. Mol Cell Biol. 1989 Mar;9(3):875–884. doi: 10.1128/mcb.9.3.875. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Kemphues K. J., Raff R. A., Kaufman T. C., Raff E. C. Mutation in a structural gene for a beta-tubulin specific to testis in Drosophila melanogaster. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3991–3995. doi: 10.1073/pnas.76.8.3991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Linder P., Lasko P. F., Ashburner M., Leroy P., Nielsen P. J., Nishi K., Schnier J., Slonimski P. P. Birth of the D-E-A-D box. Nature. 1989 Jan 12;337(6203):121–122. doi: 10.1038/337121a0. [DOI] [PubMed] [Google Scholar]
  10. Ma L., Siemssen E. D., Noteborn H. M., van der Eb A. J. The xeroderma pigmentosum group B protein ERCC3 produced in the baculovirus system exhibits DNA helicase activity. Nucleic Acids Res. 1994 Oct 11;22(20):4095–4102. doi: 10.1093/nar/22.20.4095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Ma L., Westbroek A., Jochemsen A. G., Weeda G., Bosch A., Bootsma D., Hoeijmakers J. H., van der Eb A. J. Mutational analysis of ERCC3, which is involved in DNA repair and transcription initiation: identification of domains essential for the DNA repair function. Mol Cell Biol. 1994 Jun;14(6):4126–4134. doi: 10.1128/mcb.14.6.4126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Mounkes L. C., Fuller M. T. The DUG gene of Drosophila melanogaster encodes a structural and functional homolog of the S. cerevisiae SUG1 predicted ATPase associated with the 26S proteasome. Gene. 1998 Jan 12;206(2):165–174. doi: 10.1016/s0378-1119(97)00564-7. [DOI] [PubMed] [Google Scholar]
  13. Mounkes L. C., Jones R. S., Liang B. C., Gelbart W., Fuller M. T. A Drosophila model for xeroderma pigmentosum and Cockayne's syndrome: haywire encodes the fly homolog of ERCC3, a human excision repair gene. Cell. 1992 Dec 11;71(6):925–937. doi: 10.1016/0092-8674(92)90389-t. [DOI] [PubMed] [Google Scholar]
  14. O'Connell P. O., Rosbash M. Sequence, structure, and codon preference of the Drosophila ribosomal protein 49 gene. Nucleic Acids Res. 1984 Jul 11;12(13):5495–5513. doi: 10.1093/nar/12.13.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Park E., Guzder S. N., Koken M. H., Jaspers-Dekker I., Weeda G., Hoeijmakers J. H., Prakash S., Prakash L. RAD25 (SSL2), the yeast homolog of the human xeroderma pigmentosum group B DNA repair gene, is essential for viability. Proc Natl Acad Sci U S A. 1992 Dec 1;89(23):11416–11420. doi: 10.1073/pnas.89.23.11416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Qiu H., Park E., Prakash L., Prakash S. The Saccharomyces cerevisiae DNA repair gene RAD25 is required for transcription by RNA polymerase II. Genes Dev. 1993 Nov;7(11):2161–2171. doi: 10.1101/gad.7.11.2161. [DOI] [PubMed] [Google Scholar]
  17. Regan C. L., Fuller M. T. Interacting genes that affect microtubule function in Drosophila melanogaster: two classes of mutation revert the failure to complement between haync2 and mutations in tubulin genes. Genetics. 1990 May;125(1):77–90. doi: 10.1093/genetics/125.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Regan C. L., Fuller M. T. Interacting genes that affect microtubule function: the nc2 allele of the haywire locus fails to complement mutations in the testis-specific beta-tubulin gene of Drosophila. Genes Dev. 1988 Jan;2(1):82–92. doi: 10.1101/gad.2.1.82. [DOI] [PubMed] [Google Scholar]
  19. Robertson H. M., Preston C. R., Phillis R. W., Johnson-Schlitz D. M., Benz W. K., Engels W. R. A stable genomic source of P element transposase in Drosophila melanogaster. Genetics. 1988 Mar;118(3):461–470. doi: 10.1093/genetics/118.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Rubin G. M., Spradling A. C. Genetic transformation of Drosophila with transposable element vectors. Science. 1982 Oct 22;218(4570):348–353. doi: 10.1126/science.6289436. [DOI] [PubMed] [Google Scholar]
  21. Schaeffer L., Roy R., Humbert S., Moncollin V., Vermeulen W., Hoeijmakers J. H., Chambon P., Egly J. M. DNA repair helicase: a component of BTF2 (TFIIH) basic transcription factor. Science. 1993 Apr 2;260(5104):58–63. doi: 10.1126/science.8465201. [DOI] [PubMed] [Google Scholar]
  22. Sweder K. S., Hanawalt P. C. The COOH terminus of suppressor of stem loop (SSL2/RAD25) in yeast is essential for overall genomic excision repair and transcription-coupled repair. J Biol Chem. 1994 Jan 21;269(3):1852–1857. [PubMed] [Google Scholar]
  23. Talesa V., Culetto E., Schirru N., Bernardi H., Fedon Y., Toutant J. P., Arpagaus M. Characterization of a null mutation in ace-1, the gene encoding class A acetylcholinesterase in the nematode Caenorhabditis elegans. FEBS Lett. 1995 Jan 9;357(3):265–268. doi: 10.1016/0014-5793(94)01343-y. [DOI] [PubMed] [Google Scholar]
  24. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vermeulen W., Scott R. J., Rodgers S., Müller H. J., Cole J., Arlett C. F., Kleijer W. J., Bootsma D., Hoeijmakers J. H., Weeda G. Clinical heterogeneity within xeroderma pigmentosum associated with mutations in the DNA repair and transcription gene ERCC3. Am J Hum Genet. 1994 Feb;54(2):191–200. [PMC free article] [PubMed] [Google Scholar]
  26. Walker J. E., Saraste M., Runswick M. J., Gay N. J. Distantly related sequences in the alpha- and beta-subunits of ATP synthase, myosin, kinases and other ATP-requiring enzymes and a common nucleotide binding fold. EMBO J. 1982;1(8):945–951. doi: 10.1002/j.1460-2075.1982.tb01276.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Weeda G., Eveno E., Donker I., Vermeulen W., Chevallier-Lagente O., Taïeb A., Stary A., Hoeijmakers J. H., Mezzina M., Sarasin A. A mutation in the XPB/ERCC3 DNA repair transcription gene, associated with trichothiodystrophy. Am J Hum Genet. 1997 Feb;60(2):320–329. [PMC free article] [PubMed] [Google Scholar]
  28. Weeda G., van Ham R. C., Vermeulen W., Bootsma D., van der Eb A. J., Hoeijmakers J. H. A presumed DNA helicase encoded by ERCC-3 is involved in the human repair disorders xeroderma pigmentosum and Cockayne's syndrome. Cell. 1990 Aug 24;62(4):777–791. doi: 10.1016/0092-8674(90)90122-u. [DOI] [PubMed] [Google Scholar]
  29. Williams L. D., Shaw B. R. Protonated base pairs explain the ambiguous pairing properties of O6-methylguanine. Proc Natl Acad Sci U S A. 1987 Apr;84(7):1779–1783. doi: 10.1073/pnas.84.7.1779. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Genetics are provided here courtesy of Oxford University Press

RESOURCES