CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.

The CRISPR-Cas (Clustered Regularly Interspaced Short Palindrome Repeats--CRISPR associated proteins) system provides adaptive immunity in archaea and bacteria. A hallmark of CRISPR-Cas is the involvement of short crRNAs that guide associated proteins in the destruction of invading DNA or RNA. We pr...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Ingeborg Scholz, Sita J Lange, Stephanie Hein, Wolfgang R Hess, Rolf Backofen
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/d96360b65aad4a368027a1a1afc42190
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:d96360b65aad4a368027a1a1afc42190
record_format	dspace
spelling	oai:doaj.org-article:d96360b65aad4a368027a1a1afc421902021-11-18T07:57:10ZCRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.1932-620310.1371/journal.pone.0056470https://doaj.org/article/d96360b65aad4a368027a1a1afc421902013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23441196/?tool=EBIhttps://doaj.org/toc/1932-6203The CRISPR-Cas (Clustered Regularly Interspaced Short Palindrome Repeats--CRISPR associated proteins) system provides adaptive immunity in archaea and bacteria. A hallmark of CRISPR-Cas is the involvement of short crRNAs that guide associated proteins in the destruction of invading DNA or RNA. We present three fundamentally distinct processing pathways in the cyanobacterium Synechocystis sp. PCC6803 for a subtype I-D (CRISPR1), and two type III systems (CRISPR2 and CRISPR3), which are located together on the plasmid pSYSA. Using high-throughput transcriptome analyses and assays of transcript accumulation we found all CRISPR loci to be highly expressed, but the individual crRNAs had profoundly varying abundances despite single transcription start sites for each array. In a computational analysis, CRISPR3 spacers with stable secondary structures displayed a greater ratio of degradation products. These structures might interfere with the loading of the crRNAs into RNP complexes, explaining the varying abundancies. The maturation of CRISPR1 and CRISPR2 transcripts depends on at least two different Cas6 proteins. Mutation of gene sll7090, encoding a Cmr2 protein led to the disappearance of all CRISPR3-derived crRNAs, providing in vivo evidence for a function of Cmr2 in the maturation, regulation of expression, Cmr complex formation or stabilization of CRISPR3 transcripts. Finally, we optimized CRISPR repeat structure prediction and the results indicate that the spacer context can influence individual repeat structures.Ingeborg ScholzSita J LangeStephanie HeinWolfgang R HessRolf BackofenPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 2, p e56470 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ingeborg Scholz Sita J Lange Stephanie Hein Wolfgang R Hess Rolf Backofen CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
description	The CRISPR-Cas (Clustered Regularly Interspaced Short Palindrome Repeats--CRISPR associated proteins) system provides adaptive immunity in archaea and bacteria. A hallmark of CRISPR-Cas is the involvement of short crRNAs that guide associated proteins in the destruction of invading DNA or RNA. We present three fundamentally distinct processing pathways in the cyanobacterium Synechocystis sp. PCC6803 for a subtype I-D (CRISPR1), and two type III systems (CRISPR2 and CRISPR3), which are located together on the plasmid pSYSA. Using high-throughput transcriptome analyses and assays of transcript accumulation we found all CRISPR loci to be highly expressed, but the individual crRNAs had profoundly varying abundances despite single transcription start sites for each array. In a computational analysis, CRISPR3 spacers with stable secondary structures displayed a greater ratio of degradation products. These structures might interfere with the loading of the crRNAs into RNP complexes, explaining the varying abundancies. The maturation of CRISPR1 and CRISPR2 transcripts depends on at least two different Cas6 proteins. Mutation of gene sll7090, encoding a Cmr2 protein led to the disappearance of all CRISPR3-derived crRNAs, providing in vivo evidence for a function of Cmr2 in the maturation, regulation of expression, Cmr complex formation or stabilization of CRISPR3 transcripts. Finally, we optimized CRISPR repeat structure prediction and the results indicate that the spacer context can influence individual repeat structures.
format	article
author	Ingeborg Scholz Sita J Lange Stephanie Hein Wolfgang R Hess Rolf Backofen
author_facet	Ingeborg Scholz Sita J Lange Stephanie Hein Wolfgang R Hess Rolf Backofen
author_sort	Ingeborg Scholz
title	CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
title_short	CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
title_full	CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
title_fullStr	CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
title_full_unstemmed	CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.
title_sort	crispr-cas systems in the cyanobacterium synechocystis sp. pcc6803 exhibit distinct processing pathways involving at least two cas6 and a cmr2 protein.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/d96360b65aad4a368027a1a1afc42190
work_keys_str_mv	AT ingeborgscholz crisprcassystemsinthecyanobacteriumsynechocystissppcc6803exhibitdistinctprocessingpathwaysinvolvingatleasttwocas6andacmr2protein AT sitajlange crisprcassystemsinthecyanobacteriumsynechocystissppcc6803exhibitdistinctprocessingpathwaysinvolvingatleasttwocas6andacmr2protein AT stephaniehein crisprcassystemsinthecyanobacteriumsynechocystissppcc6803exhibitdistinctprocessingpathwaysinvolvingatleasttwocas6andacmr2protein AT wolfgangrhess crisprcassystemsinthecyanobacteriumsynechocystissppcc6803exhibitdistinctprocessingpathwaysinvolvingatleasttwocas6andacmr2protein AT rolfbackofen crisprcassystemsinthecyanobacteriumsynechocystissppcc6803exhibitdistinctprocessingpathwaysinvolvingatleasttwocas6andacmr2protein
_version_	1718422761015083008

CRISPR-Cas systems in the cyanobacterium Synechocystis sp. PCC6803 exhibit distinct processing pathways involving at least two Cas6 and a Cmr2 protein.

Ejemplares similares