Cytoplasmic Dynein Function Is Essential in Drosophila melanogaster

Abstract

The microtubule motor cytoplasmic dynein has been implicated in a variety of intracellular transport processes. We previously identified and characterized the Drosophila gene Dhc64C, which encodes a cytoplasmic dynein heavy chain. To investigate the function of the cytoplasmic dynein motor, we initiated a mutational analysis of the Dhc64C dynein gene. A small deletion that removes the chromosomal region containing the heavy chain gene was used to isolate EMS-induced lethal mutations that define at least eight essential genes in the region. Germline transformation with a Dhc64C transgene rescued 16 mutant alleles in the single complementation group that identifies the dynein heavy chain gene. All 16 alleles were hemizygous lethal, which demonstrates that the cytoplasmic dynein heavy chain gene Dhc64C is essential for Drosophila development. Furthermore, our failure to recover somatic clones of cells homozygous for a Dhc64C mutation indicates that cytoplasmic dynein function is required for cell viability in several Drosophila tissues. The intragenic complementation of dynein alleles reveals multiple mutant phenotypes including male and/or female sterility, bristle defects, and defects in eye development.

Drosophila roadblock and Chlamydomonas LC7: A Conserved Family of Dynein-associated Proteins Involved in Axonal Transport, Flagellar Motility, and Mitosis

“…For example, previous clonal analysis of the null allele robl l(2)k10408 (before cloning of the gene) showed reduced clone size but a normal frequency of clone generation (Roch et al, 1998). However, clonal analysis of a strong cytoplasmic dynein heavy chain allele found a complete absence of clones in most flies and only a few small clones in the wing and abdomen observed in some flies (Gepner et al, 1996). Together these experiments suggest that whereas the dynein heavy chain gene is required for cell viability, cells can accomplish all minimally required cell autonomous dynein roles in the absence of robl.…”

Patterning the Drosophila embryo: A paradigm for RNA‐based developmental genetic regulation

“…Dynein, as assessed by immunostaining for the dynein heavy chain Dhc64C, accumulates into the early oocyte in a similar manner to osk mRNA (Li, McGrail, Serr, & Hays, 1994). In early egg chambers dynein forms part of a multiprotein complex that also contains Egalitarian (Egl), Bicaudal‐D (Bic‐D), and Lissencephaly‐1 (Lis‐1), and mutations affecting any dynein complex gene block oocyte differentiation (Gepner et al, 1996; Liu, Xie, & Steward, 1999; Mach & Lehmann, 1997; McGrail & Hays, 1997; Navarro, Puthalakath, Adams, Strasser, & Lehmann, 2004; Swan, Nguyen, & Suter, 1999). RNA injection experiments discussed in the preceding section examined both bcd and osk RNAs and suggested that this early stage of osk localization involves dynein‐directed transport (Clark et al, 2007).…”

The Drosophila Ninein homologue Bsg25D cooperates with Ensconsin in myonuclear positioning

“…Standard Drosophila genetics were performed at 25°C. The following stocks were used: apterous ME -NLS::dsRed (Richardson et al, 2007), ens swo (Metzger et al, 2012), Df-ens Δ3277 (Sung et al, 2008), khc 8 (Brendza et al, 1999), dhc [4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] (Gepner et al, 1996), plp 5 (Martinez-Campos et al, 2004), Dmef2-Gal4 (Ranganayakulu et al, 1998), rP298-Gal4 (Menon and Chia, 2001), ubiquitin-Gal4[3xP3-GFP] (Baena-Lopez et al, 2013, UAS-Ens-HA (Metzger et al, 2012), UAS-2xEGFP (Halfon et al, 2002), UAS-EB1-YFP (Bulgakova et al, 2013), and patronin Δ2 (gift from M. Gonzalez-Gaitan, University of Geneva, Geneva, Switzerland). w 1118 (3605), yw (1495), UAS-Rab7.GFP (42706), UASp-GFP.Golgi (30902), UAS-GFP.KDEL (9899), Df(2L)BSC693 (26545), Df(3L) BSC441 ( 24945), hs-FLP, hs-SceI (25679), γ-tub23C A6-2 (5728), and γ-tub23C A14-9 (7041) were obtained from the Bloomington Drosophila Stock Center.…”