Analysis of substrate specificity of human DHHC protein acyltransferases using a yeast expression system

Ohno, Yusuke; Kashio, Atsushi; Ogata, Ren; Ishitomi, Akihiro; Yamazaki, Yuki; Kihara, Akio

doi:10.1091/mbc.e12-05-0336

Cited by 83 publications

(77 citation statements)

References 33 publications

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“…This finding demonstrates for the first time that the DHHC1/10 (z1/11) subfamily proteins are functional palmitoylating enzymes in mammalian cells. In the final stage of our manuscript preparation, another group reported that human DHHC1 and -10 (z11) show the palmitoylating activity in the yeast system (53). Thus, this in silico approach can complement experimental approaches to clarify protein functions regulated by post-translational modifications.…”

Section: Discussionmentioning

confidence: 92%

In Silico Screening for Palmitoyl Substrates Reveals a Role for DHHC1/3/10 (zDHHC1/3/11)-mediated Neurochondrin Palmitoylation in Its Targeting to Rab5-positive Endosomes

Oku

Takahashi

Fukata

et al. 2013

Journal of Biological Chemistry

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Background:The identification of palmitoyl substrate-enzyme pairs is important for elucidating physiological roles of protein palmitoylation. Results: In silico screening revealed neurochondrin palmitoylation. DHHC1/3/10 were identified as neurochondrin palmitoylating enzymes and were essential for targeting of neurochondrin to Rab5-positive endosomes. Conclusion: Neurochondrin and the DHHC1/10 and DHHC3/7 subfamilies represent novel substrate-enzyme pairs. Significance: In silico palmitoyl screening is useful for clarifying functions of palmitoylation.

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Section: Discussionmentioning

confidence: 92%

In Silico Screening for Palmitoyl Substrates Reveals a Role for DHHC1/3/10 (zDHHC1/3/11)-mediated Neurochondrin Palmitoylation in Its Targeting to Rab5-positive Endosomes

Oku

Takahashi

Fukata

et al. 2013

Journal of Biological Chemistry

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“…Here we show that Rho2 is a substrate for Erf2 palmitoyltransferase and its accessory protein Erf4. It has been proposed that Erf2 belongs to class III DHHC proteins due to its preference for palmitoylated lipidated Ras and Rho GTPases (56). Whereas in budding yeast Erf2-Shr5 (Erf4 ortholog) mediates palmitoylation of Ras2, Rho2, and Rho3 (57), in fission yeast Ras1 and Rho3 are substrates for the Erf2-Erf4 complex (16,49).…”

Section: Discussionmentioning

confidence: 99%

Rho2 Palmitoylation Is Required for Plasma Membrane Localization and Proper Signaling to the Fission Yeast Cell Integrity Mitogen-Activated Protein Kinase Pathway

Sánchez-Mir

Franco

Martín‐García

et al. 2014

Molecular and Cellular Biology

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bThe fission yeast small GTPase Rho2 regulates morphogenesis and is an upstream activator of the cell integrity pathway, whose key element, mitogen-activated protein kinase (MAPK) Pmk1, becomes activated by multiple environmental stimuli and controls several cellular functions. Here we demonstrate that farnesylated Rho2 becomes palmitoylated in vivo at cysteine-196 within its carboxyl end and that this modification allows its specific targeting to the plasma membrane. Unlike that of other palmitoylated and prenylated GTPases, the Rho2 control of morphogenesis and Pmk1 activity is strictly dependent upon plasma membrane localization and is not found in other cellular membranes. Indeed, artificial plasma membrane targeting bypassed the Rho2 need for palmitoylation in order to signal. Detailed functional analysis of Rho2 chimeras fused to the carboxyl end from the essential GTPase Rho1 showed that GTPase palmitoylation is partially dependent on the prenylation context and confirmed that Rho2 signaling is independent of Rho GTP dissociation inhibitor (GDI) function. We further demonstrate that Rho2 is an in vivo substrate for DHHC family acyltransferase Erf2 palmitoyltransferase. Remarkably, Rho3, another Erf2 target, negatively regulates Pmk1 activity in a Rho2-independent fashion, thus revealing the existence of cross talk whereby both GTPases antagonistically modulate the activity of this MAPK cascade.

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“…Palmitoylation is thought to increase the association of substrate proteins with cellular membranes. Recent studies in yeast have identified the Asp-His-His-Cys (DHHC) family of proteins as palmitoyl acyltransferases (PATs) (Ohno et al, 2012). There are 23 evolutionarily conserved DHHC genes in mammals.…”

Section: Introductionmentioning

confidence: 99%

Palmitoylation of plakophilin is required for desmosome assembly

Roberts

Johnson

McGuinn

et al. 2014

Journal of Cell Science

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Desmosomes are prominent adhesive junctions found in various epithelial tissues. The cytoplasmic domains of desmosomal cadherins interact with a host of desmosomal plaque proteins, including plakophilins, plakoglobin and desmoplakin, which, in turn, recruit the intermediate filament cytoskeleton to sites of cell-cell contact. Although the individual components of the desmosome are known, mechanisms regulating the assembly of this junction are poorly understood. Protein palmitoylation is a posttranslational lipid modification that plays an important role in protein trafficking and function. Here, we demonstrate that multiple desmosomal components are palmitoylated in vivo. Pharmacologic inhibition of palmitoylation disrupts desmosome assembly at cell-cell borders. We mapped the site of plakophilin palmitoylation to a conserved cysteine residue present in the armadillo repeat domain. Mutation of this single cysteine residue prevents palmitoylation, disrupts plakophilin incorporation into the desmosomal plaque and prevents plakophilindependent desmosome assembly. Finally, plakophilin mutants unable to become palmitoylated act in a dominant-negative manner to disrupt proper localization of endogenous desmosome components and decrease desmosomal adhesion. Taken together, these data demonstrate that palmitoylation of desmosomal components is important for desmosome assembly and adhesion.

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Analysis of substrate specificity of human DHHC protein acyltransferases using a yeast expression system

Abstract: Twenty-one human DHHC proteins formed acyl intermediates. Seventeen of the proteins exhibited protein acyltransferase activities. DHHC1, 10, 14, and 16 are novel protein acyltransferases. DHHC proteins are classified into three classes based on substrate specificities.

Cited by 83 publications

References 33 publications

In Silico Screening for Palmitoyl Substrates Reveals a Role for DHHC1/3/10 (zDHHC1/3/11)-mediated Neurochondrin Palmitoylation in Its Targeting to Rab5-positive Endosomes

In Silico Screening for Palmitoyl Substrates Reveals a Role for DHHC1/3/10 (zDHHC1/3/11)-mediated Neurochondrin Palmitoylation in Its Targeting to Rab5-positive Endosomes

Rho2 Palmitoylation Is Required for Plasma Membrane Localization and Proper Signaling to the Fission Yeast Cell Integrity Mitogen-Activated Protein Kinase Pathway

Palmitoylation of plakophilin is required for desmosome assembly

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