The tomato subtilase family includes several cell death-related proteinases with caspase specificity

Scientific Reports
Sven ReichardtA Schaller

Abstract

Phytaspases are Asp-specific subtilisin-like plant proteases that have been likened to animal caspases with respect to their regulatory function in programmed cell death (PCD). We identified twelve putative phytaspase genes in tomato that differed widely in expression level and tissue-specific expression patterns. Most phytaspase genes are tandemly arranged on tomato chromosomes one, four, and eight, and many belong to taxon-specific clades, e.g. the P69 clade in the nightshade family, suggesting that these genes evolved by gene duplication after speciation. Five tomato phytaspases (SlPhyts) were expressed in N. benthamiana and purified to homogeneity. Substrate specificity was analyzed in a proteomics assay and with a panel of fluorogenic peptide substrates. Similar to animal caspases, SlPhyts recognized an extended sequence motif including Asp at the cleavage site. Clear differences in cleavage site preference were observed implying different substrates in vivo and, consequently, different physiological functions. A caspase-like function in PCD was confirmed for five of the seven tested phytaspases. Cell death was triggered by ectopic expression of SlPhyts 2, 3, 4, 5, 6 in tomato leaves by agro-infiltration, as well as in sta...Continue Reading

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Citations

Feb 23, 2019·Journal of Experimental Botany·Rafael Andrade BuonoMoritz K Nowack
Oct 23, 2019·Biological Chemistry·Shiyu ChenMatthew Bogyo
Jul 4, 2020·Proceedings of the National Academy of Sciences of the United States of America·Judith K PaulusRenier A L van der Hoorn
Aug 6, 2019·Frontiers in Plant Science·Svetlana V TrusovaAndrey B Vartapetian
Mar 29, 2020·Science·S ReichardtA Schaller
Dec 1, 2020·The New Phytologist·Shuaishuai WangSuomeng Dong
Jan 20, 2021·Journal of Experimental Botany·Alice Godson, Renier A L van der Hoorn

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Methods Mentioned

BETA
transgenic
size-exclusion chromatography
Size-
exclusion chromatography
transgenics
PCR
gel filtration

Software Mentioned

PICS
ITAG
GraphPad Prism
ClustalX
BLAST
GSTree
Clustal X
IceLogos
CulstalX

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