dtctex-1 , the Drosophila melanogaster homolog of a putative murine t-complex distorter encoding a dynein light chain, i
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O R I GI N A L P A P E R
C. Caggese á R. Moschetti á G. Ragone P. Barsanti á R. Caizzi
dtctex-1, the Drosophila melanogaster homolog of a putative murine t-complex distorter encoding a dynein light chain, is required for production of functional sperm Received: 9 August 2000 / Accepted: 29 November 2000 / Published online: 21 February 2001 Ó Springer-Verlag 2001
Abstract Tctex-1 is a light chain of the cytoplasmic and ¯agellar dyneins and a candidate for one of the distorter products that cause transmission ratio distortion in mice. We report the identi®cation, characterization, and a preliminary mutational analysis of the function of the Drosophila melanogaster dtctex-1 gene, the putative ortholog of the mammalian tctex-1 gene family. Four Ptransposon insertions which disrupt the 5¢ untranslated region of dtctex-1 are viable in homozygous form but cause male sterility due to the production of non-motile sperm. In males homozygous for dtctex-1 mutant alleles the dtctex-1 transcript is undetectable, while in homozygous females transcripts of lower molecular weight are present. By secondary mobilization of P-element insertions several revertants and new mutant alleles carrying deletions in the 5¢ UTR region of the gene were produced and characterized by PCR and by Northern analysis. Key words Tctex-1 á Dynein á Drosophila melanogaster á Male sterility á P-element-induced mutations
Introduction Dyneins are microtubule-associated molecular motors ®rst identi®ed as the ATPase required for the generation of bends or wave propagation in ciliary and ¯agellar axonemes (Warner et al. 1989). Moreover, dyneins are axonemal components of sperm tails, and have been
Communicated by E. K. F. Bautz C. Caggese (&) á R. Moschetti á G. Ragone P. Barsanti á R. Caizzi Dipartimento di Anatomia Patologica e di Genetica (DAPEG), Sez. Genetica, UniversitaÁ di Bari, Via Amendola 165/A, 70126 Bari, Italy E-mail: [email protected] Fax: +39-80-5443386
shown to be required for sperm motility in organisms as evolutionarily remote as sea urchins, insects and mammals (Gagnon et al. 1994; Kurek et al. 1998). Highmolecular-weight complexes related to axonemal dyneins in overall structural organization and in their component polypeptides have also been isolated from organisms and tissues that lack cilia or ¯agella. These are the cytoplasmic dyneins, whose functions are still incompletely understood but are essentially related to intracellular transport and mobility. For example, cytoplasmic dyneins are involved in organelle transport and localization (Paschal and Vallee 1987; Schroer et al. 1989; Corthesy-Theulaz et al. 1992; Xiang et al. 1994), and also play a role in chromosomal movements during cell division (Li et al. 1993; Cottingham and Hoyt 1997; Neesen et al. 1997; Inoue et al. 1998; Bowman et al. 1999). While the two heavy chains of the cytoplasmic dynein complex contain the ATPase and motor activity and interact with microtubules, intermediate and light chains are thought to be responsible for speci®c interactions with dierent
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