Functional characterization of NopT1 and NopT2, two type III effectors of Bradyrhizobium japonicum

dc.contributor.author	Fotiadis, CT	en
dc.contributor.author	Dimou, M	en
dc.contributor.author	Georgakopoulos, DG	en
dc.contributor.author	Katinakis, P	en
dc.contributor.author	Tampakaki, AP	en
dc.date.accessioned	2014-06-06T06:51:49Z
dc.date.available	2014-06-06T06:51:49Z
dc.date.issued	2012	en
dc.identifier.issn	03781097	en
dc.identifier.uri	http://dx.doi.org/10.1111/j.1574-6968.2011.02466.x	en
dc.identifier.uri	http://62.217.125.90/xmlui/handle/123456789/5716
dc.subject	Cysteine protease	en
dc.subject	Effector protein	en
dc.subject	Myristoylation	en
dc.subject	Palmitoylation	en
dc.subject	Type III secretion	en
dc.subject.other	cysteine proteinase	en
dc.subject.other	n [n (3 carboxyoxirane 2 carbonyl)leucyl]agmatine	en
dc.subject.other	NopT1 protein	en
dc.subject.other	NopT2 protein	en
dc.subject.other	unclassified drug	en
dc.subject.other	acylation	en
dc.subject.other	article	en
dc.subject.other	autoproteolysis	en
dc.subject.other	bacterial gene	en
dc.subject.other	bacterial plant disease	en
dc.subject.other	Bradyrhizobium japonicum	en
dc.subject.other	cell death	en
dc.subject.other	controlled study	en
dc.subject.other	enzyme activity	en
dc.subject.other	Escherichia coli	en
dc.subject.other	gene mutation	en
dc.subject.other	gene overexpression	en
dc.subject.other	mutational analysis	en
dc.subject.other	Nicotiana benthamiana	en
dc.subject.other	nonhuman	en
dc.subject.other	NopT1 gene	en
dc.subject.other	NopT2 gene	en
dc.subject.other	plant cell	en
dc.subject.other	plant leaf	en
dc.subject.other	priority journal	en
dc.subject.other	protein cleavage	en
dc.subject.other	protein degradation	en
dc.subject.other	protein expression	en
dc.subject.other	symptom	en
dc.subject.other	tobacco	en
dc.subject.other	transient expression	en
dc.subject.other	type III secretion system	en
dc.subject.other	Amino Acid Motifs	en
dc.subject.other	Amino Acid Sequence	en
dc.subject.other	Bacterial Proteins	en
dc.subject.other	Bradyrhizobium	en
dc.subject.other	Cysteine Endopeptidases	en
dc.subject.other	Molecular Sequence Data	en
dc.subject.other	Tobacco	en
dc.subject.other	Bacteria (microorganisms)	en
dc.subject.other	Bradyrhizobium japonicum	en
dc.subject.other	Escherichia coli	en
dc.subject.other	Nicotiana obtusifolia	en
dc.subject.other	Nicotiana tabacum	en
dc.title	Functional characterization of NopT1 and NopT2, two type III effectors of Bradyrhizobium japonicum	en
heal.type	journalArticle	en
heal.identifier.primary	10.1111/j.1574-6968.2011.02466.x	en
heal.publicationDate	2012	en
heal.abstract	NopT1 and NopT2, putative type III effectors from the plant symbiotic bacterium Bradyrhizobium japonicum, are predicted to belong to a family of YopT/AvrPphB effectors, which are cysteine proteases. In the present study, we showed that both NopT1 and NopT2 indeed possess cysteine protease activity. When overexpressed in Escherichia coli, both NopT1 and NopT2 undergo autoproteolytic processing which is largely abolished in the presence of E-64, a papain family-specific inhibitor. Mutations of NopT1 disrupting either the catalytic triad or the putative autoproteolytic site reduce or markedly abolish the protease activity. Autocleavage likely occurs between residues K48 and M49, though another potential cleavage site is also possible. NopT1 also elicitis HR-like cell death when transiently expressed in tobacco plants and its cysteine protease activity is essential for this ability. In contrast, no macroscopic symptoms were observed for NopT2. Furthermore, mutational analysis provided evidence that NopT1 may undergo acylation inside plant cells and that this would be required for its capacity to elicit HR-like cell death in tobacco. © 2012 Federation of European Microbiological Societies.	en
heal.journalName	FEMS Microbiology Letters	en
dc.identifier.issue	1	en
dc.identifier.volume	327	en
dc.identifier.doi	10.1111/j.1574-6968.2011.02466.x	en
dc.identifier.spage	66	en
dc.identifier.epage	77	en