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. 1997 May 15;16(10):2671–2681. doi: 10.1093/emboj/16.10.2671

The NTPase/helicase activities of Drosophila maleless, an essential factor in dosage compensation.

C G Lee 1, K A Chang 1, M I Kuroda 1, J Hurwitz 1
PMCID: PMC1169878  PMID: 9184214

Abstract

Drosophila maleless (mle) is required for X chromosome dosage compensation and is essential for male viability. Maleless protein (MLE) is highly homologous to human RNA helicase A and the bovine counterpart of RNA helicase A, nuclear helicase II. In this report, we demonstrate that MLE protein, overexpressed and purified from Sf9 cells infected with recombinant baculovirus, possesses RNA/DNA helicase, adenosine triphosphatase (ATPase) and single-stranded (ss) RNA/ssDNA binding activities, properties identical to RNA helicase A. Using site-directed mutagenesis, we created a mutant of MLE (mle-GET) that contains a glutamic acid in place of lysine in the conserved ATP binding site A. In vitro biochemical analysis showed that this mutation abolished both NTPase and helicase activities of MLE but affected the ability of MLE to bind to ssRNA, ssDNA and guanosine triphosphate (GTP) less severely. In vivo, mle-GET protein could still localize to the male X chromosome coincidentally with the male-specific lethal-1 protein, MSL-1, but failed to complement mle1 mutant males. These results indicate that the NTPase/helicase activities are essential functions of MLE for dosage compensation, perhaps utilized for chromatin remodeling of X-linked genes.

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Selected References

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

  1. Baker B. S., Gorman M., Marín I. Dosage compensation in Drosophila. Annu Rev Genet. 1994;28:491–521. doi: 10.1146/annurev.ge.28.120194.002423. [DOI] [PubMed] [Google Scholar]
  2. Ballabio A., Willard H. F. Mammalian X-chromosome inactivation and the XIST gene. Curr Opin Genet Dev. 1992 Jun;2(3):439–447. doi: 10.1016/s0959-437x(05)80155-8. [DOI] [PubMed] [Google Scholar]
  3. Bashaw G. J., Baker B. S. The msl-2 dosage compensation gene of Drosophila encodes a putative DNA-binding protein whose expression is sex specifically regulated by Sex-lethal. Development. 1995 Oct;121(10):3245–3258. doi: 10.1242/dev.121.10.3245. [DOI] [PubMed] [Google Scholar]
  4. Bell L. R., Horabin J. I., Schedl P., Cline T. W. Positive autoregulation of sex-lethal by alternative splicing maintains the female determined state in Drosophila. Cell. 1991 Apr 19;65(2):229–239. doi: 10.1016/0092-8674(91)90157-t. [DOI] [PubMed] [Google Scholar]
  5. Bone J. R., Lavender J., Richman R., Palmer M. J., Turner B. M., Kuroda M. I. Acetylated histone H4 on the male X chromosome is associated with dosage compensation in Drosophila. Genes Dev. 1994 Jan;8(1):96–104. doi: 10.1101/gad.8.1.96. [DOI] [PubMed] [Google Scholar]
  6. Cline T. W. Two closely linked mutations in Drosophila melanogaster that are lethal to opposite sexes and interact with daughterless. Genetics. 1978 Dec;90(4):683–698. doi: 10.1093/genetics/90.4.683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Côté J., Quinn J., Workman J. L., Peterson C. L. Stimulation of GAL4 derivative binding to nucleosomal DNA by the yeast SWI/SNF complex. Science. 1994 Jul 1;265(5168):53–60. doi: 10.1126/science.8016655. [DOI] [PubMed] [Google Scholar]
  8. Franke A., DeCamillis M., Zink D., Cheng N., Brock H. W., Paro R. Polycomb and polyhomeotic are constituents of a multimeric protein complex in chromatin of Drosophila melanogaster. EMBO J. 1992 Aug;11(8):2941–2950. doi: 10.1002/j.1460-2075.1992.tb05364.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Fukunaga A., Tanaka A., Oishi K. Maleless, a recessive autosomal mutant of Drosophila melanogaster that specifically kills male zygotes. Genetics. 1975 Sep;81(1):135–141. doi: 10.1093/genetics/81.1.135. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gibson T. J., Thompson J. D. Detection of dsRNA-binding domains in RNA helicase A and Drosophila maleless: implications for monomeric RNA helicases. Nucleic Acids Res. 1994 Jul 11;22(13):2552–2556. doi: 10.1093/nar/22.13.2552. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. 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]
  12. Gorman M., Franke A., Baker B. S. Molecular characterization of the male-specific lethal-3 gene and investigations of the regulation of dosage compensation in Drosophila. Development. 1995 Feb;121(2):463–475. doi: 10.1242/dev.121.2.463. [DOI] [PubMed] [Google Scholar]
  13. Gorman M., Kuroda M. I., Baker B. S. Regulation of the sex-specific binding of the maleless dosage compensation protein to the male X chromosome in Drosophila. Cell. 1993 Jan 15;72(1):39–49. doi: 10.1016/0092-8674(93)90048-u. [DOI] [PubMed] [Google Scholar]
  14. Kuroda M. I., Kernan M. J., Kreber R., Ganetzky B., Baker B. S. The maleless protein associates with the X chromosome to regulate dosage compensation in Drosophila. Cell. 1991 Sep 6;66(5):935–947. doi: 10.1016/0092-8674(91)90439-6. [DOI] [PubMed] [Google Scholar]
  15. Lee C. G., Hurwitz J. A new RNA helicase isolated from HeLa cells that catalytically translocates in the 3' to 5' direction. J Biol Chem. 1992 Mar 5;267(7):4398–4407. [PubMed] [Google Scholar]
  16. Lee C. G., Hurwitz J. Human RNA helicase A is homologous to the maleless protein of Drosophila. J Biol Chem. 1993 Aug 5;268(22):16822–16830. [PubMed] [Google Scholar]
  17. Lee D. Y., Hayes J. J., Pruss D., Wolffe A. P. A positive role for histone acetylation in transcription factor access to nucleosomal DNA. Cell. 1993 Jan 15;72(1):73–84. doi: 10.1016/0092-8674(93)90051-q. [DOI] [PubMed] [Google Scholar]
  18. Maroni G., Plaut W. Dosage compensation in Drosophila melanogaster triploids. I. Autoradiographic study. Chromosoma. 1973;40(4):361–377. doi: 10.1007/BF00399428. [DOI] [PubMed] [Google Scholar]
  19. Meyer B. J., Casson L. P. Caenorhabditis elegans compensates for the difference in X chromosome dosage between the sexes by regulating transcript levels. Cell. 1986 Dec 26;47(6):871–881. doi: 10.1016/0092-8674(86)90802-0. [DOI] [PubMed] [Google Scholar]
  20. Mukherjee A. S., Beermann W. Synthesis of ribonucleic acid by the X-chromosomes of Drosophila melanogaster and the problem of dosage compensation. Nature. 1965 Aug 14;207(998):785–786. doi: 10.1038/207785a0. [DOI] [PubMed] [Google Scholar]
  21. Pause A., Méthot N., Sonenberg N. The HRIGRXXR region of the DEAD box RNA helicase eukaryotic translation initiation factor 4A is required for RNA binding and ATP hydrolysis. Mol Cell Biol. 1993 Nov;13(11):6789–6798. doi: 10.1128/mcb.13.11.6789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Peterson C. L., Herskowitz I. Characterization of the yeast SWI1, SWI2, and SWI3 genes, which encode a global activator of transcription. Cell. 1992 Feb 7;68(3):573–583. doi: 10.1016/0092-8674(92)90192-f. [DOI] [PubMed] [Google Scholar]
  23. Pirrotta V., Bickel S., Mariani C. Developmental expression of the Drosophila zeste gene and localization of zeste protein on polytene chromosomes. Genes Dev. 1988 Dec;2(12B):1839–1850. doi: 10.1101/gad.2.12b.1839. [DOI] [PubMed] [Google Scholar]
  24. Richter L., Bone J. R., Kuroda M. I. RNA-dependent association of the Drosophila maleless protein with the male X chromosome. Genes Cells. 1996 Mar;1(3):325–336. doi: 10.1046/j.1365-2443.1996.26027.x. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. 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]
  27. Saurin A. J., Borden K. L., Boddy M. N., Freemont P. S. Does this have a familiar RING? Trends Biochem Sci. 1996 Jun;21(6):208–214. [PubMed] [Google Scholar]
  28. Spradling A. C., Rubin G. M. Transposition of cloned P elements into Drosophila germ line chromosomes. Science. 1982 Oct 22;218(4570):341–347. doi: 10.1126/science.6289435. [DOI] [PubMed] [Google Scholar]
  29. Tsukiyama T., Daniel C., Tamkun J., Wu C. ISWI, a member of the SWI2/SNF2 ATPase family, encodes the 140 kDa subunit of the nucleosome remodeling factor. Cell. 1995 Dec 15;83(6):1021–1026. doi: 10.1016/0092-8674(95)90217-1. [DOI] [PubMed] [Google Scholar]
  30. Tsukiyama T., Wu C. Purification and properties of an ATP-dependent nucleosome remodeling factor. Cell. 1995 Dec 15;83(6):1011–1020. doi: 10.1016/0092-8674(95)90216-3. [DOI] [PubMed] [Google Scholar]
  31. Turner B. M., Birley A. J., Lavender J. Histone H4 isoforms acetylated at specific lysine residues define individual chromosomes and chromatin domains in Drosophila polytene nuclei. Cell. 1992 Apr 17;69(2):375–384. doi: 10.1016/0092-8674(92)90417-b. [DOI] [PubMed] [Google Scholar]
  32. Wassarman D. A., Steitz J. A. RNA splicing. Alive with DEAD proteins. Nature. 1991 Feb 7;349(6309):463–464. doi: 10.1038/349463a0. [DOI] [PubMed] [Google Scholar]
  33. Wolffe A. P., Pruss D. Targeting chromatin disruption: Transcription regulators that acetylate histones. Cell. 1996 Mar 22;84(6):817–819. doi: 10.1016/s0092-8674(00)81059-4. [DOI] [PubMed] [Google Scholar]
  34. Zhang S., Grosse F. Nuclear DNA helicase II unwinds both DNA and RNA. Biochemistry. 1994 Apr 5;33(13):3906–3912. doi: 10.1021/bi00179a016. [DOI] [PubMed] [Google Scholar]
  35. Zhang S., Maacke H., Grosse F. Molecular cloning of the gene encoding nuclear DNA helicase II. A bovine homologue of human RNA helicase A and Drosophila Mle protein. J Biol Chem. 1995 Jul 7;270(27):16422–16427. doi: 10.1074/jbc.270.27.16422. [DOI] [PubMed] [Google Scholar]

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