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Summer Research Fellowship Programme of India's Science Academies

CHARACTERIZATION OF PREFOLDIN ANTIBODIES

Shree Nithya N

VIT, Near Katpadi Road, Vellore, Tamil Nadu 632014

Dr. Vimlesh Kumar

Indian Institute of Science Education and Research Bhopal, Bhopal Bypass Road, Bhauri, Bhopal, Madhya Pradesh 462066

Abstract

Prefoldin (PFD) is a heterohexameric complex functioning as a molecular chaperone, a class of proteins assisting in the ATP- dependant correct folding of various other proteins. Prefoldin is known to capture an unfolded actin or tubulin and specifically transfer it to cytosolic chaperonin (c-cpn), in eukaryotes. Having the appearance of a jellyfish, prefoldin has 6 subunits ; the body of PFD comprises of a double barrel assembly with six long coiled coil protrusions, resembling tentacles. In order to understand the localization of this protein, Prefoldin, specifically in the neuromuscular junction, Drosophila is used as a model. The Drosophila neuromuscular junction is a well-established model for studying fundamental questions about synaptic development, plasticity and function. Many molecules involved in synaptic transmission are said to be conserved between Drosophila and vertebrates, and hence the results of the study can be extrapolated to vertebrates. The neuromuscular junction in the A2 hemisegment of the 3rd instar larva of Drosophila is studied. Confirmation of the location of the occurrence of the protein is carried out by immunocytochemical assays and blotting techniques along with a series of genetic experiments involving RNAi investigation via the GAL4 system. The adoption of the bipartite GAL4 system in the field of fly genetics has resulted in driving transgene expression in a precise spatiotemporal pattern. The aim of this study involves the characterization of specific antibodies raised against Prefoldin subunit 5 and subunit 6, and the localization of these proteins in the neuromuscular junction of Drosophila, along with genetic knockdown studies using RNAi experiments.

Keywords: prefoldin, chaperone, neuromuscular junction, RNAi

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