title: AtFACE-2, a functional prenylated protein protease from Arabidopsis thaliana related to mammalian Ras-converting enzymes.
abstract: Eukaryotic proteins containing a CAAX (A is aliphatic amino acid) C-terminal tetrapeptide sequence generally undergo a lipid modification, the addition of a prenyl group. Proteins that are modified by prenylation, such as Ras GTPases, can be subsequently modified by a proteolytic event that removes a C-terminal tripeptide (AAX). Two distinct proteases have been identified that are involved in the CAAX proteolytic step, FACE-1/Ste24 and FACE-2/Rce1. These proteases have different enzymatic properties, substrate specificities, and biological functions. However, a proposal has been made that plants lack a FACE-2/Rce1-type protease. Here, we describe the isolation of a cDNA from Arabidopsis thaliana that encodes a 311-aa protein with characteristics that are similar to the FACE-2/Rce1 group of enzymes. Northern blot analysis demonstrates widespread expression of this gene in plant tissues. Heterologous expression of the A. thaliana cDNA in yeast restores CAAX proteolytic activity to yeast lacking native CAAX proteases. The recombinant protein produced in this system displays an in vivo substrate specificity profile distinct from AtSte24 and cleaves a farnesylated CAAX tetrapeptide in vitro. These results provide evidence for the existence of a previously unsuspected plant FACE-2/Rce1 ortholog and support the evolutionary conservation of dual CAAX proteolytic systems in eukaryotes.
authors.affiliation: Departamento de Bioquímica y Biología Molecular, Instituto Universitario de Oncología, Universidad de Oviedo, Campus del Christo, 33006-Oviedo, Spain.
authors.forename: Juan
authors.initials: J
authors.lastname: Cadiñanos
authors.forename: Ignacio
authors.initials: I
authors.lastname: Varela
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Mandel
authors.forename: Walter K
authors.initials: WK
authors.lastname: Schmidt
authors.forename: Araceli
authors.initials: A
authors.lastname: Díaz-Perales
authors.forename: Carlos
authors.initials: C
authors.lastname: López-Otín
authors.forename: José M P
authors.initials: JM
authors.lastname: Freije
mesh_terms: D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000737:chemistry; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000302:isolation & purification; D010450:Endopeptidases; D008565:Membrane Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism; D043484:Proprotein Convertases; D017368:Protein Prenylation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000494:pharmacology; D029701:Saccharomyces cerevisiae Proteins; D013379:Substrate Specificity; D015003:Yeasts / Q000378:metabolism
chemical_list: D029681:Arabidopsis Proteins; D018076:DNA, Complementary; D008565:Membrane Proteins; D009842:Oligopeptides; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D010450:Endopeptidases; D043484:Proprotein Convertases; C105193:RCE1 protein, S cerevisiae; D008666:Metalloendopeptidases; C104079:STE24 protein, Arabidopsis