The Ras Target AF-6 is a Substrate of the Fam Deubiquitinating Enzyme

Taya, Shinichiro; Yamamoto, Takaharu; Kanô, Kyoko; Kawano, Yoji; Iwamatsu, Akihiro; Tsuchiya, Tomoko; Tanaka, Keiji; Kanai‐Azuma, Masami; Wood, Stephen A.; Mattick, John S.; Kaibuchi, Kozo

doi:10.1083/jcb.142.4.1053

Cited by 105 publications

(101 citation statements)

References 48 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Immunoblot experiments using the testicular and ovarian extracts clearly demonstrated that anti-Af-6 and anti-Usp9x Ab specifically recognized a specific single band of approximately 205 and 220 kDa respectively. These data clearly confirmed the high specificity of each Ab as was demonstrated in previous studies (Taya et al 1998, Kanai-Azuma et al 2000. PMSG treatment revealed an enhancement of Usp9x expression level, slightly in the testes and strongly in the ovaries (right-hand plates in Fig.…”

Section: Localization Of Af-6 and Usp9x In Sertoli Cellssupporting

confidence: 76%

“…As well as the post-translational regulation of tight and adherens junction proteins, a gap junction protein, Connexin43, is also degraded via the ubiquitin proteasome pathway (Laing & Beyer 1995, Laing et al 1997. Moreover, our previous report (Taya et al 1998) showed a possible regulation of Af-6, an integral component of tight and adherens junctions, by an Usp9x-mediated ubiquitin pathway. These studies therefore clearly indicate that the balance between positive and negative regulators of ubiquitylation may likely be involved in the junction dynamics in various epithelial cells, including Sertoli and granulosa cells of mammalian gonads.…”

Section: Discussionmentioning

confidence: 99%

“…Most interestingly, Af-6 has been shown to be one substrate of Usp9x in MDCK cells (Taya et al 1998), although no appreciable genetic interaction was found between Usp9x ( faf) and Af-6 (canoe) homologues in Drosophila eye development (Chen et al 2000). In MDCK cells, Af-6 interacts with Usp9x both in vivo and in vitro, Af-6 is ubiquitylated in intact cells and Usp9x prevents the ubiquitylation of Af-6 in this cell line.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

A close correlation in the expression patterns of Af-6 and Usp9x in Sertoli and granulosa cells of mouse testis and ovary

Satô¹,

Kanai²,

Noma³

et al. 2004

Reproduction

Self Cite

View full text Add to dashboard Cite

Usp9x, an X-linked deubiquitylating enzyme, is stage dependently expressed in the supporting cells (i.e. Sertoli cells and granulosa cells) and germ cells during mouse gametogenesis. Af-6, a cell junction protein, has been identified as a substrate of Usp9x, suggesting a possible association between Usp9x and Af-6 in spermatogenesis and oogenesis. In this study, we examined the expression pattern of Af-6 and Usp9x and their intracellular localization in testes and ovaries of mice treated with or without pregnant mare serum gonadotropin (PMSG), an FSH-like hormone. In both testes and ovaries, Af-6 expression was predominantly observed in supporting cells, as well as in steroidogenic cells, but not in any germ cells. In Sertoli cells, Af-6 was continuously expressed throughout postnatal and adult stages, where both Af-6 and Usp9x were enriched at the sites of Sertoli-Sertoli and Sertoli -spermatid junctions especially at stages XI -VI. In the granulosa cells, Af-6, as well as Usp9x, was highly expressed in primordial and primary follicles, but its expression rapidly decreased after the late-secondary follicle stage. Interestingly, in PMSG-treated mice, the expression levels of Af-6 and Usp9x were synchronously enhanced, slightly in Sertoli cells and strongly in granulosa cells of the late-secondary and Graafian follicles. Such closely correlated expression patterns between Af-6 and Usp9x clearly suggest that Af-6 may be deubiquitylated by Usp9x in both Sertoli and granulosa cells. It further suggests that the post-translational regulation of Af-6 by Usp9x may be one potential pathway to control the cell adhesion dynamics in mammalian gametogenesis.

show abstract

Section: Localization Of Af-6 and Usp9x In Sertoli Cellssupporting

confidence: 76%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

A close correlation in the expression patterns of Af-6 and Usp9x in Sertoli and granulosa cells of mouse testis and ovary

Satô¹,

Kanai²,

Noma³

et al. 2004

Reproduction

Self Cite

View full text Add to dashboard Cite

show abstract

“…1 A and B). Among the putative ERG-binding proteins (SI Appendix, Table S1) identified by both pulldown and immunoprecipitation was a deubiquitinase enzyme, USP9X (15). We validated the physical interaction of ERG with USP9X by immunoblot analysis of coimmunoprecipitation with endogenous ERG (Fig.…”

Section: Resultsmentioning

confidence: 99%

Ablation of the oncogenic transcription factor ERG by deubiquitinase inhibition in prostate cancer

Wang

Kollipara

Srivastava

et al. 2014

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

112

115

View full text Add to dashboard Cite

The transcription factor E-twenty-six related gene (ERG), which is overexpressed through gene fusion with the androgen-responsive gene transmembrane protease, serine 2 (TMPRSS2) in ∼40% of prostate tumors, is a key driver of prostate carcinogenesis. Ablation of ERG would disrupt a key oncogenic transcriptional circuit and could be a promising therapeutic strategy for prostate cancer treatment. Here, we show that ubiquitin-specific peptidase 9, X-linked (USP9X), a deubiquitinase enzyme, binds ERG in VCaP prostate cancer cells expressing TMPRSS2-ERG and deubiquitinates ERG in vitro. USP9X knockdown resulted in increased levels of ubiquitinated ERG and was coupled with depletion of ERG. Treatment with the USP9X inhibitor WP1130 resulted in ERG degradation both in vivo and in vitro, impaired the expression of genes enriched in ERG and prostate cancer relevant gene signatures in microarray analyses, and inhibited growth of ERG-positive tumors in three mouse xenograft models. Thus, we identified USP9X as a potential therapeutic target in prostate cancer cells and established WP1130 as a lead compound for the development of ERGdepleting drugs. P rostate cancer is the most common malignancy in men and the second or third leading cause of male cancer-related death in most Western countries, including the United States (1). Advanced prostate cancer initially responds to androgen ablation therapy, but hormone-refractory prostate cancer often times recurs, which has limited treatment options. Fusions of E-twentysix (ETS) transcription factor genes with androgen-responsive genes (2), mainly transmembrane protease, serine 2 (TMPRSS2), are present in up to 80% of prostate cancers. Patients with the most common ETS gene fusion TMPRSS2-ETS related gene (TMPRSS2-ERG) have a higher incidence of metastatic disease and cancer-related death compared with fusion-negative patients (3, 4), and in castration-resistant prostate cancer, TMPRSS2-ERG expression is frequently reactivated (5). In support of ERG being a key driver of prostate cancer, depletion of ERG by RNAi decreases proliferation and/or invasiveness in prostate cancer cell lines (2, 6), and ectopic expression of ERG in transgenic mice was shown to promote prostate oncogenesis in cooperation with the loss of tumor suppressors (7-12). TMPRSS2-driven overexpression of ERG controls a transcriptional network related to the development of prostate cancer and its progression to metastatic disease (13,14). This crucial role of ERG and the high incidence of the TMPRSS2-ERG gene fusion in prostate cancer have catapulted this protein into the forefront of new targets for therapeutic intervention (3,8). In the present study, we report the discovery of a deubiquitinase that stabilizes ERG in prostate cancer cells and demonstrate that pharmacological inhibition of this enzyme causes ERG depletion. Results USP9X is an ERG-Binding Protein.Proteins that interact with ERG in prostate cancer cells may modulate its activity, localization, or stability and could be harnessed as therapeutic target...

show abstract

“…It has two splice variants, l-afadin and s-afadin, but only the larger splice variant (l-afadin) can bind to actin (Mandai et al, 1997). s-Afadin (also known as AF-6), on the other hand, can localize to both the plasma membrane and nucleus (Buchert et al, 2007) and has also been reported to form a complex with and serve as a substrate for Fam [a deubiquitinating enzyme (Taya et al, 1998)]. This implies that proteolytic degradation of AF-6 occurs via the ubiquitin-proteasome pathway.…”

Section: A Cell-cell Actin-based Adherens Junctionsmentioning

confidence: 99%