Xenopus Egg Extracts as a Simplified Model System for Structure–Function Studies of Dynein Regulators

Żyłkiewicz, Eliza; Stukenberg, P. Todd

doi:10.1007/978-1-4939-0329-0_8

Cited by 3 publications

(3 citation statements)

References 23 publications

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“…The utility of the Xenopus extract system to study a variety of biological problems is clear (e.g., (Murray, 1991;Nikos, 2012;Zyłkiewicz and Stukenberg, 2014) and many others). Here we are focusing on its use in the study of proteolysis, where the simplicity of the ubiquitylation and degradation assays that the extract system allows has been key to the significant advances that have resulted (see (Klotzbucher et al, 2002;Salic et al, 2000;Vosper et al, 2009) for examples of Ub protocols).…”

Section: Xenopus Extract Systemsmentioning

confidence: 99%

Ubiquitin-mediated proteolysis in Xenopus extract

McDowell¹,

Philpott²

2016

Int. J. Dev. Biol.

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The small protein modifier, ubiquitin, can be covalently attached to proteins in the process of ubiquitylation, resulting in a variety of functional outcomes. In particular, the most commonly-associated and well-studied fate for proteins modified with ubiquitin is their ultimate destruction: degradation by the 26S proteasome via the ubiquitin-proteasome system, or digestion in lysosomes by proteolytic enzymes. From the earliest days of ubiquitylation research, a reliable and versatile "cell-in-a-test-tube" system has been employed in the form of cytoplasmic extracts from the eggs and embryos of the frog Xenopus laevis. Biochemical studies of ubiquitin and protein degradation using this system have led to significant advances particularly in the study of ubiquitin-mediated proteolysis, while the versatility of Xenopus as a developmental model has allowed investigation of the in vivo consequences of ubiquitylation. Here we describe the use and history of Xenopus extract in the study of ubiquitin-mediated protein degradation, and highlight the versatility of this system that has been exploited to uncover mechanisms and consequences of ubiquitylation and proteolysis.

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Section: Xenopus Extract Systemsmentioning

confidence: 99%

Ubiquitin-mediated proteolysis in Xenopus extract

McDowell¹,

Philpott²

2016

Int. J. Dev. Biol.

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“…The utility of the Xenopus extract system to study a variety of biological problems is clear (e.g., [15,28,29] and many others). Here we are focusing on its use in the study of proteolysis, where the simplicity of the ubiquitylation and degradation assays that the extract system allows has been key to the significant advances that have resulted (see [30,31,32] for examples of Ub protocols).…”

Section: Xenopus Extract Systemsmentioning

confidence: 99%

Ubiquitin-mediated proteolysis inXenopusextract

McDowell

Philpott

2016

Preprint

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The small protein modifier, ubiquitin, can be covalently attached to proteins in the process of ubiquitylation, resulting in a variety of functional outcomes. In particular, the most commonly-associated and well-studied fate for proteins modified with ubiquitin is their ultimate destruction: degradation by the 26S proteasome via the ubiquitin-proteasome system, or digestion in lysosomes by proteolytic enzymes.From the earliest days of ubiquitylation research, a reliable and versatile "cell-in-a-test-tube" system has been employed in the form of cytoplasmic extracts from the eggs and embryos of the frog Xenopus laevis. Biochemical studies of ubiquitin and protein degradation using this system have led to significant advances particularly in the study of ubiquitin-mediated proteolysis, while the versatility of Xenopus as a developmental model has allowed investigation of the in vivo consequences of ubiquitylation. Here we describe the use and history of Xenopus extract in the study of ubiquitin-mediated protein degradation, and highlight the versatility of this system that has been exploited to uncover mechanisms and consequences of ubiquitylation and proteolysis.

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“…Large volumes of extracts can be prepared from eggs and oocytes by centrifugation, and these contain vast arrays of proteins that enable reconstitution of cell cycle events, nuclear transportation, microtubule polymerisation and apoptosis (e.g. Blow and Laskey, 1986 , Laskey et al, 1978 , Leno et al, 1992 , Nutt, 2012 , Zylkiewicz and Stukenberg, 2014 , Deming and Kornbluth, 2006 ). Moreover, the cell-free nature of the extract system means the molecular machinery involved in these processes can be probed by manipulation of extracts with immune-depletion or neutralisation of an endogenous protein of interest, followed by subsequent rescue by addition of recombinant protein (e.g.…”

Section: Modelling With Xenopus Eggs and Cleavage mentioning

confidence: 99%

An oncologist׳s friend: How Xenopus contributes to cancer research

Hardwick

Philpott

2015

Developmental Biology

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One of the most striking features of the Xenopus system is the versatility in providing a unique range of both in vitro and in vivo models that are rapid, accessible and easily manipulated. Here we present an overview of the diverse contribution that Xenopus has made to advance our understanding of tumour biology and behaviour; a contribution that goes beyond the traditional view of Xenopus as a developmental model organism. From the utility of the egg and oocyte extract system to the use of whole embryos as developmental or induced tumour models, the Xenopus system has been fundamental to investigation of cell cycle mechanisms, cell metabolism, cell signalling and cell behaviour, and has allowed an increasing appreciation of the parallels between early development and the pathogenesis of tumour progression and metastasis. Although not the prototypical oncological model system, we propose that Xenopus is an adaptable and multifunctional tool in the oncologist׳s arsenal.

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Xenopus Egg Extracts as a Simplified Model System for Structure–Function Studies of Dynein Regulators

Cited by 3 publications

References 23 publications

Ubiquitin-mediated proteolysis in Xenopus extract

Ubiquitin-mediated proteolysis in Xenopus extract

Ubiquitin-mediated proteolysis inXenopusextract

An oncologist׳s friend: How Xenopus contributes to cancer research

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