Cloning and characterization of AWP1, a novel protein that associates with serine/threonine kinase PRK1 in vivo

Duan, Wei; Sun, Bingang; Li, Tian Wei; Tan, Bee Jen; Lee, Mui Khin; Teo, Tian Seng

doi:10.1016/s0378-1119(00)00365-6

Cited by 43 publications

(45 citation statements)

References 30 publications

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“…The proteins were transferred onto Protran membranes (Schleicher & Schuell). After blocking with 5% nonfat milk in 20 mM Tris-HCl, pH 7.5, 0.15 M NaCl, and 0.05% Tween 20 at room temperature for 1 h, the blots were probed with primary antibodies as described (16). The immunoblots were then analyzed using the SuperSignal West Dura chemiluminescence detection kit (Pierce) according to the manufacturer's recommendation.…”

Section: Methodsmentioning

confidence: 99%

The Last 10 Amino Acid Residues beyond the Hydrophobic Motif Are Critical for the Catalytic Competence and Function of Protein Kinase Cα

Yeong

Zhu

Smith

et al. 2006

Journal of Biological Chemistry

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The segment C-terminal to the hydrophobic motif at the V5 domain of protein kinase C (PKC) is the least conserved both in length and in amino acid identity among all PKC isozymes. By generating serial truncation mutants followed by biochemical and functional analyses, we show here that the very C terminus of PKC␣ is critical in conferring the full catalytic competence to the kinase and for transducing signals in cells. Deletion of one C-terminal amino acid residue caused the loss of ϳ60% of the catalytic activity of the mutant PKC␣, whereas deletion of 10 C-terminal amino acid residues abrogated the catalytic activity of PKC␣ in immune complex kinase assays. The PKC␣ C-terminal truncation mutants were found to lose their ability to activate mitogen-activated protein kinase, to rescue apoptosis induced by the inhibition of endogenous PKC in COS cells, and to augment melatonin-stimulated neurite outgrowth. Furthermore, molecular dynamics simulations revealed that the deletion of 1 or 10 C-terminal residues results in the deformation of the V5 domain and the ATP-binding pocket, respectively. Finally, PKC␣ immunoprecipitated using an antibody against its C terminus had only marginal catalytic activity compared with that of the PKC␣ immunoprecipitated by an antibody against its N terminus. Therefore, the very C-terminal tail of PKC␣ is a novel determinant of the catalytic activity of PKC and a promising target for selective modulation of PKC␣ function. Molecules that bind preferentially to the very C terminus of distinct PKC isozymes and suppress their catalytic activity may constitute a new class of selective inhibitors of PKC.

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Section: Methodsmentioning

confidence: 99%

The Last 10 Amino Acid Residues beyond the Hydrophobic Motif Are Critical for the Catalytic Competence and Function of Protein Kinase Cα

Yeong

Zhu

Smith

et al. 2006

Journal of Biological Chemistry

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“…AF140722). The database search using the amino acid sequence as query showed homology to several zinc-finger proteins including the human and mouse PRK1-associated protein AWP1 (22), PVPR3 (23), human and mouse zinc-finger proteins ZNF216 (24), Xenopus ubiquitin-like fusion proteins (25), and the ascidian posterior end mark (PEM6) protein (26). All these proteins showed homology to OSISAP1 in the AN1-type zincfinger region, which is present at the C terminus of the protein, stretching between amino acids 101 and 163.…”

Section: Analysis Of Osisap1mentioning

confidence: 99%

Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco

Mukhopadhyay

Vij

Tyagi

2004

Proc. Natl. Acad. Sci. U.S.A.

439

339

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Stress perception and signal transduction leading to tolerance involve a complex interplay of different gene products. We describe here the isolation and characterization of an intronless gene (OSISAP1) from rice encoding a zinc-finger protein that is induced after different types of stresses, namely cold, desiccation, salt, submergence, and heavy metals as well as injury. The gene is also induced by stress hormone abscisic acid. Overexpression of the gene in transgenic tobacco conferred tolerance to cold, dehydration, and salt stress at the seed-germination/seedling stage as reflected by the percentage of germination/green seedlings, the fresh weight of seedlings, and their developmental pattern. Thus, OSISAP1 seems to be an important determinant of stress response in plants.

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“…Although AWP1 has been shown to be expressed in mouse and human early embryos as well as in several human tissues (Duan et al, 2000), its physiological function remains unknown.…”

Section: Discussionmentioning

confidence: 99%

“…AWP1 PRK1, a lipid-activated kinase belonging to the protein kinase C superfamily (Duan et al, 2000). AWP1 contains A20 and AN1 (Fenner et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

“…Recently, AWP1 has also been shown to et al, 2011) and protein export during biogenesis of the peroxisome, a subcellular organelle (Miyata et al, 2012). AWP1 is expressed in several human tissues and also in the early stages of mammalian development (Duan et al, 2000), but its physiological function remains unknown. We have found that AWP1 affects the expression of NPB and NC markers in the neural fold by mediating Wnt/ -catenin signaling.…”

Section: Introductionmentioning

confidence: 99%

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Essential role of AWP1 in neural crest specification in Xenopus

Seo¹,

Park²,

Hong³

et al. 2013

Int. J. Dev. Biol.

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The neural crest (NC) comprises a transient and multipotent embryonic cell population, which gives rise to a wide variety of cell types, including craniofacial cartilage, melanocytes, and neurons and glia of the peripheral nervous system. The NC is induced by the integrated action of Wnt, FGF, and BMP signaling, and its cell fates are subsequently specified by a genetic cascade of specific transcription factors. Here we describe a critical role of AWP1 in NC induction during Xenopus early development. Xenopus AWP1 (XAWP1) was found to be expressed in the presumptive preplacodal ectoderm, neural tissue, and posterior dorsal mesoderm, but was absent in the neural fold along the anterior-posterior axis of the neurulae. Notably, XAWP1 was induced by FGF8a in naïve ectodermal tissue. XAWP1-depleted embryos exhibited defects in pigmentation, craniofacial cartilage, and in the dorsal fin. A knockdown of XAWP1 impaired both endogenous and the FGF8a or Wnt8-induced expression of NC markers without affecting mesoderm formation. Furthermore, NC induction inhibited by XAWP1 depletion was rescued by co-expression of activating forms of -catenin or TCF3. In addition, overexpression of XAWP1, in concert with BMP inhibition, induced the expression of neural plate border specifiers, Pax3 and Msx1, and these regulatory factors recovered NC induction in the XAWP1-depleted embryos. -catenin stability and Wnt-responsive reporter activity were also impaired in AWP1-depleted cells. Taken together, these results suggest that XAWP1 functions as a mediator of Wnt signaling to regulate NC specification.

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Cloning and characterization of AWP1, a novel protein that associates with serine/threonine kinase PRK1 in vivo

Cited by 43 publications

References 30 publications

The Last 10 Amino Acid Residues beyond the Hydrophobic Motif Are Critical for the Catalytic Competence and Function of Protein Kinase Cα

The Last 10 Amino Acid Residues beyond the Hydrophobic Motif Are Critical for the Catalytic Competence and Function of Protein Kinase Cα

Overexpression of a zinc-finger protein gene from rice confers tolerance to cold, dehydration, and salt stress in transgenic tobacco

Essential role of AWP1 in neural crest specification in Xenopus

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