Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.

Transformation is a complex process that involves several interactions from the binding and uptake of naked DNA to homologous recombination. Some actions affect transformation favourably whereas others act to limit it. Here, meticulous manipulation of a single type of transforming DNA allowed for qu...

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Autores principales:	Ole Herman Ambur, Stephan A Frye, Mariann Nilsen, Eirik Hovland, Tone Tønjum
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/a91df29f1c4c4e1dbd7a029b443332b2
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spelling	oai:doaj.org-article:a91df29f1c4c4e1dbd7a029b443332b22021-11-18T07:13:33ZRestriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.1932-620310.1371/journal.pone.0039742https://doaj.org/article/a91df29f1c4c4e1dbd7a029b443332b22012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/22768309/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Transformation is a complex process that involves several interactions from the binding and uptake of naked DNA to homologous recombination. Some actions affect transformation favourably whereas others act to limit it. Here, meticulous manipulation of a single type of transforming DNA allowed for quantifying the impact of three different mediators of meningococcal transformation: NlaIV restriction, homologous recombination and the DNA Uptake Sequence (DUS). In the wildtype, an inverse relationship between the transformation frequency and the number of NlaIV restriction sites in DNA was observed when the transforming DNA harboured a heterologous region for selection (ermC) but not when the transforming DNA was homologous with only a single nucleotide heterology. The influence of homologous sequence in transforming DNA was further studied using plasmids with a small interruption or larger deletions in the recombinogenic region and these alterations were found to impair transformation frequency. In contrast, a particularly potent positive driver of DNA uptake in Neisseria sp. are short DUS in the transforming DNA. However, the molecular mechanism(s) responsible for DUS specificity remains unknown. Increasing the number of DUS in the transforming DNA was here shown to exert a positive effect on transformation. Furthermore, an influence of variable placement of DUS relative to the homologous region in the donor DNA was documented for the first time. No effect of altering the orientation of DUS was observed. These observations suggest that DUS is important at an early stage in the recognition of DNA, but does not exclude the existence of more than one level of DUS specificity in the sequence of events that constitute transformation. New knowledge on the positive and negative drivers of transformation may in a larger perspective illuminate both the mechanisms and the evolutionary role(s) of one of the most conserved mechanisms in nature: homologous recombination.Ole Herman AmburStephan A FryeMariann NilsenEirik HovlandTone TønjumPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 7, p e39742 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Ole Herman Ambur Stephan A Frye Mariann Nilsen Eirik Hovland Tone Tønjum Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.
description	Transformation is a complex process that involves several interactions from the binding and uptake of naked DNA to homologous recombination. Some actions affect transformation favourably whereas others act to limit it. Here, meticulous manipulation of a single type of transforming DNA allowed for quantifying the impact of three different mediators of meningococcal transformation: NlaIV restriction, homologous recombination and the DNA Uptake Sequence (DUS). In the wildtype, an inverse relationship between the transformation frequency and the number of NlaIV restriction sites in DNA was observed when the transforming DNA harboured a heterologous region for selection (ermC) but not when the transforming DNA was homologous with only a single nucleotide heterology. The influence of homologous sequence in transforming DNA was further studied using plasmids with a small interruption or larger deletions in the recombinogenic region and these alterations were found to impair transformation frequency. In contrast, a particularly potent positive driver of DNA uptake in Neisseria sp. are short DUS in the transforming DNA. However, the molecular mechanism(s) responsible for DUS specificity remains unknown. Increasing the number of DUS in the transforming DNA was here shown to exert a positive effect on transformation. Furthermore, an influence of variable placement of DUS relative to the homologous region in the donor DNA was documented for the first time. No effect of altering the orientation of DUS was observed. These observations suggest that DUS is important at an early stage in the recognition of DNA, but does not exclude the existence of more than one level of DUS specificity in the sequence of events that constitute transformation. New knowledge on the positive and negative drivers of transformation may in a larger perspective illuminate both the mechanisms and the evolutionary role(s) of one of the most conserved mechanisms in nature: homologous recombination.
format	article
author	Ole Herman Ambur Stephan A Frye Mariann Nilsen Eirik Hovland Tone Tønjum
author_facet	Ole Herman Ambur Stephan A Frye Mariann Nilsen Eirik Hovland Tone Tønjum
author_sort	Ole Herman Ambur
title	Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.
title_short	Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.
title_full	Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.
title_fullStr	Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.
title_full_unstemmed	Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.
title_sort	restriction and sequence alterations affect dna uptake sequence-dependent transformation in neisseria meningitidis.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/a91df29f1c4c4e1dbd7a029b443332b2
work_keys_str_mv	AT olehermanambur restrictionandsequencealterationsaffectdnauptakesequencedependenttransformationinneisseriameningitidis AT stephanafrye restrictionandsequencealterationsaffectdnauptakesequencedependenttransformationinneisseriameningitidis AT mariannnilsen restrictionandsequencealterationsaffectdnauptakesequencedependenttransformationinneisseriameningitidis AT eirikhovland restrictionandsequencealterationsaffectdnauptakesequencedependenttransformationinneisseriameningitidis AT tonetønjum restrictionandsequencealterationsaffectdnauptakesequencedependenttransformationinneisseriameningitidis
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Restriction and sequence alterations affect DNA uptake sequence-dependent transformation in Neisseria meningitidis.

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