AtFACE-2, a functional Prenylated Protein Protease from Arabidopsis thaliana Related to Mammalian Ras-converting Enzymes

Cadiñanos, Juan; Varela, Ignacio; Mandel, Daniel; Schmidt, W.; Díaz‐Perales, Araceli; López-Otı́n, Carlos; Freije, José M.P.

doi:10.1074/jbc.m306700200

Cited by 47 publications

(41 citation statements)

References 39 publications

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“…1A), indicating that a-factor processing by neither plant CaaX proteases was as efficient as by yeast Ste24. These data corroborated functional analysis of AtRCE1 function in vitro (Cadinanos et al, 2003).…”

Section: Arabidopsis Atrce1 Is Functionally Conservedsupporting

confidence: 76%

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Functional Analysis of Arabidopsis Postprenylation CaaX Processing Enzymes and Their Function in Subcellular Protein Targeting

et al. 2008

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Prenylation is a posttranslational protein modification essential for developmental processes and response to abscisic acid. Following prenylation, the three C-terminal residues are proteoliticaly removed and in turn the free carboxyl group of the isoprenyl cysteine is methylated. The proteolysis and methylation, collectively referred to as CaaX processing, are catalyzed by Ste24 endoprotease or Rce1 endoprotease and by an isoprenyl cysteine methyltransferase (ICMT). Arabidopsis (Arabidopsis thaliana) contains single STE24 and RCE1 and two ICMT homologs. Here we show that in yeast (Saccharomyces cerevisiae) AtRCE1 promoted a-mating factor secretion and membrane localization of a ROP GTPase. Furthermore, green fluorescent protein fusion proteins of AtSTE24, AtRCE1, AtICMTA, and AtICMTB are colocalized in the endoplasmic reticulum, indicating that prenylated proteins reach this compartment and that CaaX processing is likely required for subcellular targeting. AtICMTB can process yeast a-factor more efficiently than AtICMTA. Sequence and mutational analyses revealed that the higher activity AtICMTB is conferred by five residues, which are conserved between yeast Ste14p, human ICMT, and AtICMTB but not in AtICMTA. Quantitative real-time reverse transcription-polymerase chain reaction and microarray data show that AtICMTA expression is significantly lower compared to AtICMTB. AtICMTA null mutants have a wild-type phenotype, indicating that its function is redundant. However, AtICMT RNAi lines had fasciated inflorescence stems, altered phylotaxis, and developed multiple buds without stem elongation. The phenotype of the ICMT RNAi lines is similar to farnesyltransferase b-subunit mutant enhanced response to abscisic acid2 but is more subtle. Collectively, the data suggest that AtICMTB is likely the major ICMT and that methylation modulates activity of prenylated proteins.Prenylation is a posttranslational protein modification essential for the function of diverse proteins. Plant mutants lacking prenyltransferase function have pleotropic phenotypes including enlargement of the shoot apical meristem, hypersensitivity to abscisic acid (ABA), retarded growth rate, and delayed flowering (Cutler et al

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Section: Arabidopsis Atrce1 Is Functionally Conservedsupporting

confidence: 76%

“…In Arabidopsis (Arabidopsis thaliana), CaaX proteolysis is catalyzed by conserved AtSTE24 and AtRCE1/ AtFACE-2 Cadinanos et al, 2003). AtSTE24 was demonstrated to be localized in the ER .…”

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confidence: 99%

Functional Analysis of Arabidopsis Postprenylation CaaX Processing Enzymes and Their Function in Subcellular Protein Targeting

et al. 2008

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“…This can be explained by processing of lipidated G␥ subunits on the ER and Golgi, since the enzymes that remove the three C-terminal amino acids and methylate the modified cysteine are found to be integral parts of these membranes (Bracha et al, 2002;Cadinanos et al, 2003;Rodriguez-Concepcion et al, 2000). In addition, the localization on the Golgi, makes it less likely that the fusion proteins are incorrectly folded.…”

Section: Discussionmentioning

confidence: 99%

Plant G protein heterotrimers require dual lipidation motifs of Gα and Gγ and do not dissociate upon activation

Adjobo‐Hermans

Goedhart

Gadella

2006

Journal of Cell Science

111

117

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In plants one bona fide Gα subunit has been identified, as well as a single Gβ and two Gγ subunits. To study the roles of lipidation motifs in the regulation of subcellular location and heterotrimer formation in living plant cells, GFP-tagged versions of the Arabidopsis thaliana heterotrimeric G protein subunits were constructed. Mutational analysis showed that the Arabidopsis Gα subunit, GPα1, contains two lipidation motifs that were essential for plasma membrane localization. The Arabidopsis Gβ subunit, AGβ1, and the Gγ subunit, AGG1, were dependent upon each other for tethering to the plasma membrane. The second Gγ subunit, AGG2, did not require AGβ1 for localization to the plasma membrane. Like AGG1, AGG2 contains two putative lipidation motifs, both of which were necessary for membrane localization. Interaction between the subunits was studied using fluorescence resonance energy transfer (FRET) imaging by means of fluorescence lifetime imaging microscopy (FLIM). The results suggest that AGβ1 and AGG1 or AGβ1 and AGG2 can form heterodimers independent of lipidation. In addition, FLIM-FRET revealed the existence of GPα1-AGβ1-AGG1 heterotrimers at the plasma membrane. Importantly, rendering GPα1 constitutively active did not cause a FRET decrease in the heterotrimer, suggesting no dissociation upon GPα1 activation.

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“…First, the aaX portion of the CaaX motif is proteolytically removed by specific CaaX proteases (STE24 and FACE-2 in Arabidopsis) (Boyartchuk et al, 1997;Bracha et al, 2002;Cadiñ anos et al, 2003); second, the isoprenylcysteine at The transfer of a farnesyl group from farnesyl diphosphate to a CaaX protein is followed by postisoprenylation processing (proteolysis and reversible methylation). Degradation of the farnesylated protein generates FC, which is oxidized by FC lyase to farnesal.…”

Section: Introductionmentioning

confidence: 99%

Isoprenylcysteine Methylation and Demethylation Regulate Abscisic Acid Signaling inArabidopsis

et al. 2008

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Isoprenylated proteins bear an isoprenylcysteine methyl ester at the C terminus. Although isoprenylated proteins have been implicated in meristem development and negative regulation of abscisic acid (ABA) signaling, the functional role of the terminal methyl group has not been described. Here, we show that transgenic Arabidopsis thaliana plants overproducing isoprenylcysteine methyltransferase (ICMT) exhibit ABA insensitivity in stomatal closure and seed germination assays, establishing ICMT as a negative regulator of ABA signaling. By contrast, transgenic plants overproducing isoprenylcysteine methylesterase (ICME) exhibit ABA hypersensitivity in stomatal closure and seed germination assays. Thus, ICME is a positive regulator of ABA signaling. To test the hypothesis that ABA signaling is under feedback regulation at the level of isoprenylcysteine methylation, we examined the effect of ABA on ICMT and ICME gene expression. Interestingly, ABA induces ICME gene expression, establishing a positive feedback loop whereby ABA promotes ABA responsiveness of plant cells via induction of ICME expression, which presumably results in the demethylation and inactivation of isoprenylated negative regulators of ABA signaling. These results suggest strategies for metabolic engineering of crop species for drought tolerance by targeted alterations in isoprenylcysteine methylation.

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AtFACE-2, a functional Prenylated Protein Protease from Arabidopsis thaliana Related to Mammalian Ras-converting Enzymes

Cited by 47 publications

References 39 publications

Functional Analysis of Arabidopsis Postprenylation CaaX Processing Enzymes and Their Function in Subcellular Protein Targeting

Functional Analysis of Arabidopsis Postprenylation CaaX Processing Enzymes and Their Function in Subcellular Protein Targeting

Plant G protein heterotrimers require dual lipidation motifs of Gα and Gγ and do not dissociate upon activation

Isoprenylcysteine Methylation and Demethylation Regulate Abscisic Acid Signaling inArabidopsis

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