The beta 3-tubulin gene of Drosophila melanogaster is essential for viability and fertility.
We have previously shown that the beta 3-tubulin gene of Drosophila melanogaster encodes a divergent isoform expressed in a complex developmental pattern. The beta 3 gene is transiently expressed in the embryo and again in the pupa at high levels in the developing musculature, and at lower levels in several different pupal tissues of ectodermal origin. Adult expression is confined to specific somatic cells in the gonads. In some of the cell types in which it is expressed, beta 3 is the sole or predominant beta-tubulin, while in others the beta 3 protein is a minor component of the beta-tubulin pool. The sites and timing of beta 3 expression demonstrated that beta 3-tubulin is utilized primarily in cytoplasmic microtubule arrays involved in changes in cell shape and tissue organization, and suggested to us that this isoform may be functionally specialized. To determine whether the expression of the beta 3 gene is essential for normal development, and to examine the specific functions of this divergent isoform, we have generated mutations within the gene. We determined that the small deficiency Df(2R)Px2, which deletes the 60C5,6-60D9,10 region of chromosome 2, removes all of the beta 3 coding sequences, and that the distal breakpoint of the deficiency is approximately 2 kb upstream from the start of transcription of the beta 3 gene. We have generated a total of 31 ethyl methanesulfonate- or diepoxybutane-induced recessive lethal or visible mutations which map within the deficiency. These mutations define 12 new lethal complementation groups, which together with two previously identified visible mutations, altogether identify 14 genes in this interval of the second chromosome. A lethal complementation group comprising mutations in the beta 3-tubulin gene (beta Tub60D) was identified by rescue of their lethality by a wild-type copy of the gene introduced into the genome via P element-mediated germ line transformation. Analysis of the homozygous and transheterozygous phenotypes of the five beta 3 mutations recovered (alleles designated B3t1-B3t5) demonstrates that beta 3-tubulin is essential for viability and fertility.
Kimble
M
M
Department of Biology, Indiana University, Bloomington 47405.
Dettman
R W
RW
Raff
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EC
D000818:Animals; D015139:Blotting, Southern; D002874:Chromosome Mapping; D004331:Drosophila melanogaster / Q000235:genetics* / Q000254:growth & development / Q000502:physiology; D004852:Epoxy Compounds / Q000494:pharmacology; D005020:Ethyl Methanesulfonate / Q000494:pharmacology; D005260:Female; D005786:Gene Expression Regulation; D005796:Genes; D005816:Genetic Complementation Test; D007124:Immunoenzyme Techniques; D008297:Male; D009153:Mutagens; D009154:Mutation; D010641:Phenotype; D012098:Reproduction / Q000235:genetics; D015183:Restriction Mapping; D014404:Tubulin / Q000235:genetics*
D004852:Epoxy Compounds; D009153:Mutagens; D014404:Tubulin; C007366:diepoxybutane; D005020:Ethyl Methanesulfonate