Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub

Summary: Functional tags are ubiquitous in cell biology, and for studies of one chromosomal locus, the centromere, tags have been remarkably useful. The centromere directs chromosome inheritance at cell division. The location of the centromere is defined by a histone H3 variant, CENP-A. The regulati...

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Autores principales:	Catalina Salinas-Luypaert, Praveen Kumar Allu, Glennis A. Logsdon, Jennine M. Dawicki-McKenna, Craig W. Gambogi, Daniele Fachinetti, Ben E. Black
Formato:	article
Lenguaje:	EN
Publicado:	Elsevier 2021
Materias:	centromere genome editing CENP-A histone variant mitosis functional protein tags Biology (General) QH301-705.5
Acceso en línea:	https://doaj.org/article/c312cc62e4fe4719a23103d6d0993732
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spelling	oai:doaj.org-article:c312cc62e4fe4719a23103d6d09937322021-11-04T04:29:13ZGene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub2211-124710.1016/j.celrep.2021.109924https://doaj.org/article/c312cc62e4fe4719a23103d6d09937322021-11-01T00:00:00Zhttp://www.sciencedirect.com/science/article/pii/S2211124721013978https://doaj.org/toc/2211-1247Summary: Functional tags are ubiquitous in cell biology, and for studies of one chromosomal locus, the centromere, tags have been remarkably useful. The centromere directs chromosome inheritance at cell division. The location of the centromere is defined by a histone H3 variant, CENP-A. The regulation of the chromatin assembly pathway essential for centromere inheritance and function includes posttranslational modification (PTM) of key components, including CENP-A itself. Others have recently called into question the use of functional tags, with the claim that at least two widely used tags obscured the essentiality of one particular PTM, CENP-AK124 ubiquitination (ub). Here, we employ three independent gene replacement strategies that eliminate large, lysine-containing tags to interrogate these claims. Using these approaches, we find no evidence to support an essential function of CENP-AK124ub. Our general methodology will be useful to validate discoveries permitted by powerful functional tagging schemes at the centromere and other cellular locations.Catalina Salinas-LuypaertPraveen Kumar AlluGlennis A. LogsdonJennine M. Dawicki-McKennaCraig W. GambogiDaniele FachinettiBen E. BlackElsevierarticlecentromeregenome editingCENP-Ahistone variantmitosisfunctional protein tagsBiology (General)QH301-705.5ENCell Reports, Vol 37, Iss 5, Pp 109924- (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	centromere genome editing CENP-A histone variant mitosis functional protein tags Biology (General) QH301-705.5
spellingShingle	centromere genome editing CENP-A histone variant mitosis functional protein tags Biology (General) QH301-705.5 Catalina Salinas-Luypaert Praveen Kumar Allu Glennis A. Logsdon Jennine M. Dawicki-McKenna Craig W. Gambogi Daniele Fachinetti Ben E. Black Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub
description	Summary: Functional tags are ubiquitous in cell biology, and for studies of one chromosomal locus, the centromere, tags have been remarkably useful. The centromere directs chromosome inheritance at cell division. The location of the centromere is defined by a histone H3 variant, CENP-A. The regulation of the chromatin assembly pathway essential for centromere inheritance and function includes posttranslational modification (PTM) of key components, including CENP-A itself. Others have recently called into question the use of functional tags, with the claim that at least two widely used tags obscured the essentiality of one particular PTM, CENP-AK124 ubiquitination (ub). Here, we employ three independent gene replacement strategies that eliminate large, lysine-containing tags to interrogate these claims. Using these approaches, we find no evidence to support an essential function of CENP-AK124ub. Our general methodology will be useful to validate discoveries permitted by powerful functional tagging schemes at the centromere and other cellular locations.
format	article
author	Catalina Salinas-Luypaert Praveen Kumar Allu Glennis A. Logsdon Jennine M. Dawicki-McKenna Craig W. Gambogi Daniele Fachinetti Ben E. Black
author_facet	Catalina Salinas-Luypaert Praveen Kumar Allu Glennis A. Logsdon Jennine M. Dawicki-McKenna Craig W. Gambogi Daniele Fachinetti Ben E. Black
author_sort	Catalina Salinas-Luypaert
title	Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub
title_short	Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub
title_full	Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub
title_fullStr	Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub
title_full_unstemmed	Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub
title_sort	gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for cenp-ak124ub
publisher	Elsevier
publishDate	2021
url	https://doaj.org/article/c312cc62e4fe4719a23103d6d0993732
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Gene replacement strategies validate the use of functional tags on centromeric chromatin and invalidate an essential role for CENP-AK124ub

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