Drosophila myosin V has a strikingly different motor mechanism from that of vertebrate myosin Va and it is a non-processive, ensemble motor. We have independently generated two polyclonal antibody against myosin V. One polyclonal antibody was generated against the coil-coiled domain of myosin V and another polyclonal against the tail end of myosin V. Immunostaining of the Drosophila larval salivary gland localized both antibodies to the nuclear envelope. It colocalized with Drosophila lamin C, a marker for the inner membrane of the larval nuclear envelope. Immunoprecipitation experiments of larval extracts using lamin C antibody pulled down myosin V. Mutants of myosin V and over expression of myosin V in the larval salivary gland using the UAS-GAL4 system showed aberrant lamin C staining on the nuclear envelope. Immunoprecipitation experiments using myosin V antibody pulled down Rab 5 protein. Rab 5 is a small GTPase which has been shown to localize to the early endosomes. Immunostaining using Rab 5 antibody in the larval salivary gland showed staining in the nuclear envelope, similar to the staining of lamin C and myosin V. In order to determine if the localization of myosin V and Rab 5 is in the inner or outer membrane of the larval nuclear envelope, we are performing immunogold electron microscopy experiments. We postulate that Drosophila myosin V, Rab 5 and lamin C are interacting with each other to maintain the structural integrity of the nuclear envelope of the larval salivary gland.

Project Start
Project End
Budget Start
Budget End
Support Year
16
Fiscal Year
2012
Total Cost
$129,147
Indirect Cost
Name
National Heart, Lung, and Blood Institute
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Type
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