The NTPase/helicase activities of Drosophila maleless, an essential factor in dosage compensation Journal Article
| Authors: | Lee, C. G.; Chang, K. A.; Kuroda, M. I.; Hurwitz, J. |
| Article Title: | The NTPase/helicase activities of Drosophila maleless, an essential factor in dosage compensation |
| 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. |
| Keywords: | controlled study; dna binding protein; dna-binding proteins; nonhuman; mutant protein; animal cell; animals; chromosomal proteins, non-histone; amino acid substitution; protein dna binding; protein binding; drosophila; enzyme activity; acid anhydride hydrolases; structure activity relation; animalia; transcription factors; rna binding protein; rna; gene expression regulation; molecular sequence data; recombinant proteins; recombinant protein; x chromosome; base sequence; binding site; spodoptera; dosage compensation, genetic; mutagenesis, site-directed; binding sites; helicase; gene dosage; adenosine triphosphate; single stranded dna; adenosine triphosphatase; enzyme structure; site directed mutagenesis; guanosine triphosphate; rna helicase; drosophila proteins; rna helicases; enzyme localization; dna helicases; fluorescent antibody technique, indirect; genetic complementation; protein rna binding; bovinae; animals, genetically modified; baculoviridae; nucleoside-triphosphatase; rna nucleotidyltransferases; unidentified baculovirus; male; priority journal; article; dosage compensation; mle; ntpase |
| Journal Title: | EMBO Journal |
| Volume: | 16 |
| Issue: | 10 |
| ISSN: | 0261-4189 |
| Publisher: | Wiley Blackwell |
| Date Published: | 1997-05-15 |
| Start Page: | 2671 |
| End Page: | 2681 |
| Language: | English |
| DOI: | 10.1093/emboj/16.10.2671 |
| PUBMED: | 9184214 |
| PROVIDER: | scopus |
| PMCID: | PMC1169878 |
| DOI/URL: | |
| Notes: | Article -- Export Date: 17 March 2017 -- Source: Scopus |
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