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Published May 30, 1991 | public
Journal Article

Dynamin-like protein encoded by the Drosophila shibire gene associated with vesicular traffic

Abstract

Temperature-sensitive paralysis is the most striking defect of adult Drosophila carrying the shibire mutation. This is believed to be due to a reversible block of endocytosis, which prevents membrane cycling and thus depletes synaptic vesicles. The shibire mutation also affects many tissues outside the nervous system. We have now mapped and characterized the shibire gene. A 275-kilobase yeast artificial chromosome was subcloned into cosmids, among which the gene was then located by analysing with restriction-fragment length polymorphisms. A 15-kilobase fragment of wild-type DNA rescues the mutant phenotype and the sequence of two mutant alleles show differences with wild type, demonstrating that we have isolated the shibire gene. The gene encodes a protein that is highly similar to rat dynamin, 69% of the amino-acid sequence is identical. Dynamin is a GTP-driven mechanochemical enzyme related to mammalian mx-proteins and to the yeast vpsl gene product. Because the shibire gene product and dynamin have extensive similarity, we propose that they are cognate homologues. Dynamin causes microtubules to slide along each other in vitro and in extracts it is associated with a distinct, but so far uncharacterized, membrane fraction. In light of the shibire phenotype, we suggest that these proteins provide the motor for vesicular transport during endocytosis.

Additional Information

© 1991 Nature Publishing Group. Received 5 March; accepted 15 April 1991. We thank our laboratory colleagues and P. J. Johnson for advice and for reading the manuscript, E. Chang for technical assistance, B. A. Hamilton for the cDNA library, I. M. Duncan for the YAC, K. Vijay Raghavan, G. Payne and A. L Katzen for probes, the Drosophila stock center (Bloomington, Indiana) for fly strains, and C. Bazinet for making us aware of the proximity of the clathrin gene. A.M.v.d.B. was supported by EMBO and the Human Frontier Science Program. This work was supported by an NIH Program Project (to E.M.M.). The shibire nucleotide sequence has been deposited with EMBL under accession numbers X59435 and X59436.

Additional details

Created:
August 20, 2023
Modified:
October 23, 2023