DNA binding properties of the small cascade subunit Csa5.

CRISPR-Cas systems provide immunity against viral attacks in archaeal and bacterial cells. Type I systems employ a Cas protein complex termed Cascade, which utilizes small CRISPR RNAs to detect and degrade the exogenic DNA. A small sequence motif, the PAM, marks the foreign substrates. Previously, a...

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Autores principales:	Michael Daume, André Plagens, Lennart Randau
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/3a03f9be8a554109a41c8727a27fc7e6
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spelling	oai:doaj.org-article:3a03f9be8a554109a41c8727a27fc7e62021-11-25T06:03:36ZDNA binding properties of the small cascade subunit Csa5.1932-620310.1371/journal.pone.0105716https://doaj.org/article/3a03f9be8a554109a41c8727a27fc7e62014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/25148031/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203CRISPR-Cas systems provide immunity against viral attacks in archaeal and bacterial cells. Type I systems employ a Cas protein complex termed Cascade, which utilizes small CRISPR RNAs to detect and degrade the exogenic DNA. A small sequence motif, the PAM, marks the foreign substrates. Previously, a recombinant type I-A Cascade complex from the archaeon Thermoproteus tenax was shown to target and degrade DNA in vitro, dependent on a native PAM sequence. Here, we present the biochemical analysis of the small subunit, Csa5, of this Cascade complex. T. tenax Csa5 preferentially bound ssDNA and mutants that showed decreased ssDNA-binding and reduced Cascade-mediated DNA cleavage were identified. Csa5 oligomerization prevented DNA binding. Specific recognition of the PAM sequence was not observed. Phylogenetic analyses identified Csa5 as a universal member of type I-A systems and revealed three distinct groups. A potential role of Csa5 in R-loop stabilization is discussed.Michael DaumeAndré PlagensLennart RandauPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 8, p e105716 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Michael Daume André Plagens Lennart Randau DNA binding properties of the small cascade subunit Csa5.
description	CRISPR-Cas systems provide immunity against viral attacks in archaeal and bacterial cells. Type I systems employ a Cas protein complex termed Cascade, which utilizes small CRISPR RNAs to detect and degrade the exogenic DNA. A small sequence motif, the PAM, marks the foreign substrates. Previously, a recombinant type I-A Cascade complex from the archaeon Thermoproteus tenax was shown to target and degrade DNA in vitro, dependent on a native PAM sequence. Here, we present the biochemical analysis of the small subunit, Csa5, of this Cascade complex. T. tenax Csa5 preferentially bound ssDNA and mutants that showed decreased ssDNA-binding and reduced Cascade-mediated DNA cleavage were identified. Csa5 oligomerization prevented DNA binding. Specific recognition of the PAM sequence was not observed. Phylogenetic analyses identified Csa5 as a universal member of type I-A systems and revealed three distinct groups. A potential role of Csa5 in R-loop stabilization is discussed.
format	article
author	Michael Daume André Plagens Lennart Randau
author_facet	Michael Daume André Plagens Lennart Randau
author_sort	Michael Daume
title	DNA binding properties of the small cascade subunit Csa5.
title_short	DNA binding properties of the small cascade subunit Csa5.
title_full	DNA binding properties of the small cascade subunit Csa5.
title_fullStr	DNA binding properties of the small cascade subunit Csa5.
title_full_unstemmed	DNA binding properties of the small cascade subunit Csa5.
title_sort	dna binding properties of the small cascade subunit csa5.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/3a03f9be8a554109a41c8727a27fc7e6
work_keys_str_mv	AT michaeldaume dnabindingpropertiesofthesmallcascadesubunitcsa5 AT andreplagens dnabindingpropertiesofthesmallcascadesubunitcsa5 AT lennartrandau dnabindingpropertiesofthesmallcascadesubunitcsa5
_version_	1718414215415332864

DNA binding properties of the small cascade subunit Csa5.

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