The evaluation of anoxia responsive E2F DNA binding activity in the red eared slider turtle, Trachemys scripta elegans

PeerJ
Kyle K Biggar, Kenneth B Storey

Abstract

In many cases, the DNA-binding activity of a transcription factor does not change, while its transcriptional activity is greatly influenced by the make-up of bound proteins. In this study, we assessed the protein composition and DNA-binding ability of the E2F transcription factor complex to provide insight into cell cycle control in an anoxia tolerant turtle through the use of a modified ELISA protocol. This modification also permits the use of custom DNA probes that are tailored to a specific DNA binding region, introducing the ability to design capture probes for non-model organisms. Through the use of EMSA and ELISA DNA binding assays, we have successfully determined the in vitro DNA binding activity and complex dynamics of the Rb/E2F cell cycle regulatory mechanisms in an anoxic turtle, Trachemys scripta elegans. Repressive cell cycle proteins (E2F4, Rb, HDAC4 and Suv39H1) were found to significantly increase at E2F DNA-binding sites upon anoxic exposure in anoxic turtle liver. The lack of p130 involvement in the E2F DNA-bound complex indicates that anoxic turtle liver may maintain G1 arrest for the duration of stress survival.

References

Sep 29, 1999·Genes & Development·R E Kingston, G J Narlikar
Oct 26, 1999·Journal of Molecular Biology·C Muchardt, M Yaniv
Jun 23, 2001·Oncogene·R FerreiraA Harel-Bellan
Mar 9, 2002·Molecular and Cellular Biology·Geert J P L KopsBoudewijn M T Burgering
Mar 10, 2004·Biological Reviews of the Cambridge Philosophical Society·Kenneth B Storey, Janet M Storey
Sep 15, 2004·Trends in Biochemical Sciences·Adrian P BrackenKristian Helin
Apr 8, 2006·Molecular Cell·Daniele Guardavaccaro, Michele Pagano
May 10, 2007·The Journal of Experimental Biology·Kenneth B Storey, Janet M Storey
Jun 2, 2007·Cell·Patrick TrojerDanny Reinberg
Nov 7, 2007·The Journal of Cell Biology·Junjie Lu, David M Gilbert
Aug 23, 2008·Nature Reviews. Molecular Cell Biology·Sander van den Heuvel, Nicholas J Dyson
Jan 27, 2009·Current Protocols in Neuroscience·R Benjamin FreeDavid R Sibley
Oct 17, 2009·Journal of Comparative Physiology. B, Biochemical, Systemic, and Environmental Physiology·Anastasia Krivoruchko, Kenneth B Storey
Jun 2, 2010·Current Genomics·Kyle K Biggar, Kenneth B Storey
Jul 29, 2011·Journal of Experimental Zoology. Part A, Ecological Genetics and Physiology·Rabih RoufayelKenneth B Storey
Jul 12, 2013·Nature Reviews. Cancer·Subhashini Sadasivam, James A DeCaprio
Mar 16, 2017·The FEBS Journal·Zhouran WuKyle K Biggar

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Methods Mentioned

BETA
electrophoresis
enzyme-linked immunosorbent assay
co-IPs
co-IP
ELISA
electrophoretic mobility shift assay

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