290
Biotic interactions
AvrPphB proteins are required for their localization to theplasma membrane of host cells. Furthermore, thesesequences are required for bacteria expressing AvrRpm1and AvrB to elicit a resistant response by plants expressingthe cognate RPM1 protein, a plasma-membrane-associatedprotein [15]. Even more intriguing is that both the myris-toylation and palmitoylation sites of AvrRpm1 are requiredfor bacteria to be fully virulent on susceptible hosts [14].This indicates that the putative virulence target of AvrRpm1may also be associatedwith the plasma membrane.AvrPto was also shown to require N-myristoylation sites tolocalize to the host plasma membrane and trigger Pto func-tion [16]. Pto, the cognate R protein to AvrPto, also carriesputative N-myristoylation sequences, indicatingthat Ptomay be associated with the plasma membrane.Localization of Pto to the membrane has yet to be demon-strated, however, and Pto can function without theconsensus myristoylation site if driven from a strong pro-moter [17]. Additionally, a putative N-myristoylation sitehas been identified in the amino-acid sequence of RPS5, aNBS-LRR protein of
Arabidopsis
that recognizes the bac-terial Avr protein AvrPphB [18].Recognition of
P. syringae
AvrPphB has been shown tospecifically require
RPS5
and
PBS1
[19].
PBS1
has beencloned and encodes a serine-threonine kinase that appearsto be functionally distinct from Pto [20]. Interestingly, likeAvrPphB and RSP5, PBS1 also contains a putativeN-myristoylation site, potentially placing all three of theseplayers at the membrane.
Which one is my partner?
AvrBs3 family proteins of various
Xanthomonads
requirenuclear-localization sequences for transport to the nucleus,and for virulence and
R
-dependent recognition [21,22,23
•
].Thus, the corresponding R proteins might also localize tothe nucleus. The exact role of these Avr proteins in thenucleus is still unclear, but this family may be involved ininducing or repressing the expression of host genes. A tran-scriptional activation domain of AvrXa7 is required foravirulence and virulence on rice [23
•
]. When this domain issubstituted with that of the unrelated herpes simplex viralprotein16 (VP16), avirulence but not virulence activity isrestored. This result suggests that the recognition by
Xa7
mediated by the transcriptional activation domain of AvrXa7 is not sequence-specific, but rather is due to someinteraction with the transcriptional machinery of the host.The AvrXa7 activation domain is, however, required forvirulence, perhaps because it interacts with a host factor tomodify the expression of host target genes during disease.The AvrBs3 family of proteins from
Xathomonas
spp.requires the presence of NLS sequences for recognition onresistant hosts. Recent work on family member AvrBs4,however, demonstrates that its NLS sequences are dis-pensable for recognition by
Bs4
on tomato [24]. This worksuggests that recognition (interception?) by R proteins canoccur at different points along intracellular paths taken bytranslocated type-III effectors to their cellular target.Cloning and analysis of R proteins that recognize distinctAvrBs3 family members should shed light on this puzzle.The AvrBs3 proteins may be the first known case of a familyof effectors having undergone diversification aimed at a setof related host targets. Recently, several closely relatedmembers of this family (members of the AvrXa-X proteinsfrom
X. oryzae
) were mutagenized in one strain [25]. Eachmutation caused a different (and ultimately additive)impact on virulence. Moreover, none of the
avrXa-X
genestested could complement the loss of virulence functionnormally encoded by other family members. Thus, theirfunctions are non-redundant. One possible scenario is thatthese genes are evolving to not only evade host
R
func-tions, but also to maximize the number of interactions theycan have with individual members of a host protein familythat are the virulence targets. The more of these relatedtargets that can be affected the more efficient thepathogen will become.
Just holding hands or really a close dance?
The most effective R protein would be one that recognizesthe domains of its cognate Type-III effector that arerequired for virulence. In this case, effector mutations thatresult in the loss of or decreased resistance responses wouldpresumably carry a fitness cost to the pathogen. Recentmutational analyses of two homologous Type-III effectorsindicate that such ‘effective’ Rproteins might exist. The
X. campestris
Type-III effector AvrBsT is related to
Yersinia pestis
YopJ protein
.
The regions of homology include con-served sequences that encode a putative protease catalyticdomain. Directed mutations of YopJ in one of the three con-served residues required for function in related proteasesresulted in the loss of
Yersinia pestis
virulence on its normal-ly susceptible host. The same mutations in
avrBsT
resultedin the loss of recognition by the normally resistant
Nicotiania benthamiana
plants [26
•
]. Similarly,
X. campestris
pathovar (pv.)
vesicatoria
mutants were recently isolatedthat can partially overcome the pepper
Bs2
gene and alsohave reduced
avrBs2
virulence activity on
bs2
plants [27].The
avrBs2
gene was the first for which a requirement forbacterial fitness was demonstrated [28], giving rise to thenotion that
Bs2
-mediated resistance would be stable.Sequence analysis of the variant
avrBs2
genes isolated fromstrains growing on
Bs2
heterozygous plants suggests thatevolutionary pressure favors loss of avirulence but mainte-nance of virulence activities.A similar example of an ‘effective’ R protein is found inpotato. The Ry R protein recognizes the Potato VirusY(PVY) Nla protein as an avirulence protein. Nla is a pro-teinase and mutations of the active proteinase domaincompromise Ry recognition [29]. A different virus whoseproteinase has the same specificity as PVY’s, but is only63% identical to the PVY proteinase, also triggers Ry func-tion. What is not clear is whether Ry recognizes the Nla
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