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This study reports the identification and characterization of a novel gene,  Dic61B , required for male fertility in  Drosophila . Complementation mapping of a novel male sterile mutation,  ms21 , isolated in our lab revealed it to be allelic to  CG7051  at 61B1 cytogenetic region, since two  piggyBac  insertion alleles,  CG7051 c05439   and  CG7051 f07138   failed to complement.  CG7051  putatively encodes a Dynein intermediate chain. All three mutants,  ms21 ,  CG7051 c05439   and  CG7051 f07138  , exhibited absolute recessive male sterility with abnormally coiled sperm axonemes causing faulty sperm individualization as revealed by Phalloidin staining in Don Juan-GFP background. Sequencing of PCR amplicons uncovered two point mutations in  ms21  allele and confirmed the  piggyBac  insertions in  CG7051 c05439   and  CG7051 f07138   alleles to be in 5′UTR and 4 th  exon of  CG7051  respectively, excision of which reverted the male sterility.  In situ  hybridization to polytene chromosomes demonstrated  CG7051  to be a single copy gene. RT-PCR of testis RNA revealed defective splicing of the  CG7051  transcripts in mutants. Interestingly, expression of cytoplasmic dynein intermediate chain, α, β, γ tubulins and α-spectrin was normal in mutants while ultra structural studies revealed defects in the assembly of sperm axonemes. Bioinformatics further highlighted the homology of  CG7051  to axonemal dynein intermediate chain of various organisms, including DNAI1 of humans, mutations in which lead to male sterility due to immotile sperms. Based on these observations we conclude that  CG7051  encodes a novel axonemal dynein intermediate chain essential for male fertility in  Drosophila  and rename it as  Dic61B . This is the first axonemal  Dic  gene of  Drosophila  to be characterized at molecular level and shown to be required for spermatogenesis.

Drosophila Dynein Intermediate Chain Gene, Dic61B, Is Required for Spermatogenesis