Cell-specific monitoring of protein synthesis in vivo.

Analysis of general and specific protein synthesis provides important information, relevant to cellular physiology and function. However, existing methodologies, involving metabolic labelling by incorporation of radioactive amino acids into nascent polypeptides, cannot be applied to monitor protein...

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Autores principales:	Nikos Kourtis, Nektarios Tavernarakis
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2009
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/0df688c4d7ab4bec8e7f8f1c3cb7f953
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spelling	oai:doaj.org-article:0df688c4d7ab4bec8e7f8f1c3cb7f9532021-11-25T06:17:11ZCell-specific monitoring of protein synthesis in vivo.1932-620310.1371/journal.pone.0004547https://doaj.org/article/0df688c4d7ab4bec8e7f8f1c3cb7f9532009-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19234598/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Analysis of general and specific protein synthesis provides important information, relevant to cellular physiology and function. However, existing methodologies, involving metabolic labelling by incorporation of radioactive amino acids into nascent polypeptides, cannot be applied to monitor protein synthesis in specific cells or tissues, in live specimens. We have developed a novel approach for monitoring protein synthesis in specific cells or tissues, in vivo. Fluorescent reporter proteins such as GFP are expressed in specific cells and tissues of interest or throughout animals using appropriate promoters. Protein synthesis rates are assessed by following fluorescence recovery after partial photobleaching of the fluorophore at targeted sites. We evaluate the method by examining protein synthesis rates in diverse cell types of live, wild type or mRNA translation-defective Caenorhabditis elegans animals. Because it is non-invasive, our approach allows monitoring of protein synthesis in single cells or tissues with intrinsically different protein synthesis rates. Furthermore, it can be readily implemented in other organisms or cell culture systems.Nikos KourtisNektarios TavernarakisPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 4, Iss 2, p e4547 (2009)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Nikos Kourtis Nektarios Tavernarakis Cell-specific monitoring of protein synthesis in vivo.
description	Analysis of general and specific protein synthesis provides important information, relevant to cellular physiology and function. However, existing methodologies, involving metabolic labelling by incorporation of radioactive amino acids into nascent polypeptides, cannot be applied to monitor protein synthesis in specific cells or tissues, in live specimens. We have developed a novel approach for monitoring protein synthesis in specific cells or tissues, in vivo. Fluorescent reporter proteins such as GFP are expressed in specific cells and tissues of interest or throughout animals using appropriate promoters. Protein synthesis rates are assessed by following fluorescence recovery after partial photobleaching of the fluorophore at targeted sites. We evaluate the method by examining protein synthesis rates in diverse cell types of live, wild type or mRNA translation-defective Caenorhabditis elegans animals. Because it is non-invasive, our approach allows monitoring of protein synthesis in single cells or tissues with intrinsically different protein synthesis rates. Furthermore, it can be readily implemented in other organisms or cell culture systems.
format	article
author	Nikos Kourtis Nektarios Tavernarakis
author_facet	Nikos Kourtis Nektarios Tavernarakis
author_sort	Nikos Kourtis
title	Cell-specific monitoring of protein synthesis in vivo.
title_short	Cell-specific monitoring of protein synthesis in vivo.
title_full	Cell-specific monitoring of protein synthesis in vivo.
title_fullStr	Cell-specific monitoring of protein synthesis in vivo.
title_full_unstemmed	Cell-specific monitoring of protein synthesis in vivo.
title_sort	cell-specific monitoring of protein synthesis in vivo.
publisher	Public Library of Science (PLoS)
publishDate	2009
url	https://doaj.org/article/0df688c4d7ab4bec8e7f8f1c3cb7f953
work_keys_str_mv	AT nikoskourtis cellspecificmonitoringofproteinsynthesisinvivo AT nektariostavernarakis cellspecificmonitoringofproteinsynthesisinvivo
_version_	1718413966201323520

Cell-specific monitoring of protein synthesis in vivo.

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