An investigation into Mef2-interacting proteins during embryonic muscle development of Drosophila

Cepraga, Alina Anamaria 2020. An investigation into Mef2-interacting proteins during embryonic muscle development of Drosophila. PhD Thesis, Cardiff University. Item availability restricted.

Preview	PDF - Accepted Post-Print Version Download (8MB) | Preview
	PDF (Cardiff University Electronic Publication Form) - Supplemental Material Restricted to Repository staff only Download (457kB)

Abstract

The present thesis was an investigation into developing a full picture of the Mef2-interacting proteins in muscle differentiation of Drosophila melanogaster by using a mixture of wet-lab and computational approaches. Understanding how Mef2 is regulated is of general importance due to its pivotal role in cell differentiation programs. An overview of the transcription factor's interacting proteins contributes a significant part to understanding how Mef2 performs its functions. The proteins identified to interact with Mef2 at its endogenous level and in its normal pattern of expression and activity in the context of development were identified through a complex purification approach that can detect systematically protein complexes and networks. The method of choice was Tandem Affinity Purification (TAP), which offers a good recovery of the Mef2 used as bait as well as a low level of contaminants. The identified candidates were subjected to an analytical pipeline based on in silico database mining for expression and protein interactions information, as well as literature querying for existing functional information. Based on the derived information, it was possible to profile the type of proteins Mef2 interacts with during development. New candidates confirmed to be associated with muscle differentiation were shown to be quite distant towards Mef2 when known functionality based on literature was used as a relatedness coefficient. One of the identified candidates in the screen, HDAC4, was used for further biological validation in vivo. HDAC4 was extracted during a screen of 11-13 h old embryos, the developmental time point where terminal muscle differentiation occurs. The biological assessment confirmed that the two proteins Mef2 and HDAC4 interact in the context of myogenesis as HDAC4 was found to repress the expression of a Mef2 target protein in somatic muscle. The knowledge derived from this screen suggests that Mef2 does not only interact with muscle specific genes during myogenesis.

Item Type:	Thesis (PhD)
Date Type:	Completion
Status:	Unpublished
Schools:	Biosciences
Subjects:	Q Science > Q Science (General)
Date of First Compliant Deposit:	22 June 2020
Last Modified:	16 Mar 2021 02:26
URI:	https://orca.cardiff.ac.uk/id/eprint/132678

Actions (repository staff only)

Edit Item