The cloning and characterisation of alpha-adaptin in Drosophila melanogaster

The role of membrane recycling has been analysed in <I>Drosophila</I> by characterising the distribution of a clathrin coat protein throughout development. <I>Drosophila</I> α-adaptin (<I>DAA</I>) has been cloned and is well conserved with mammalian α-adaptins. It...

Full description

Bibliographic Details
Main Author: Dornan, S. C.
Published: University of Cambridge 1997
Subjects:
572
Online Access:http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.598605
Description
Summary:The role of membrane recycling has been analysed in <I>Drosophila</I> by characterising the distribution of a clathrin coat protein throughout development. <I>Drosophila</I> α-adaptin (<I>DAA</I>) has been cloned and is well conserved with mammalian α-adaptins. It is -80% similar to rat and mouse α-adaptin isoforms and a multiple sequence alignment predicts a conservation of the primary structure of the adaptin molecule. <I>DAA</I> is present as a single gene copy and maps to the left arm of chromosome 2, at 21C2.3. The hinge region of DAA has been expressed as a recombinant protein and used to generate species-specific antisera. This antisera has been used to assess the role and distribution of α-adaptin during embryogenesis. A developmental western indicates that DAA accumulates throughout embryogenesis and immunolocalisation studies of embryos using the antisera has revealed specific tissues to be highly endocytic. The neural tissues of the developing central, peripheral and stromatogastric nervous systems, cells undergoing positional rearrangements within the embryo, and the highly pinocytic garland cells are all enriched in DAA. The distribution of DAA is indicative of a role in the mediation of membrane recycling at the plasma membrane, facilitating the changing distribution of cell surface proteins required for ongoing development, and in the process of membrane remodelling within the nervous system. Immunolocalisation studies in the deficiency line, Df (2L) <I>aristaless</I> covering the chromosomal region of <I>DAA</I>, reveal that maternal DAA is present within the embryo to the late stages of embryogenesis.