The interaction of piasy with Trim32, an E3-ubiquitin ligase mutated in limb-girdle muscular dystrophy type 2H, promotes piasy degradation and regulates UVB-induced keratinocyte apoptosis through NFκB

Amador Albor, Sally El-Hizawi, Elizabeth J. Horn, Melanie Laederich, Patrick Frosk, Klaus Wrogemann, Molly Kulesz-Martin

Research output: Contribution to journalArticle

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Abstract

Protein inhibitors of activated STATs (PIAS) family members are ubiquitin-protein isopeptide ligase-small ubiquitin-like modifier ligases for diverse transcription factors. However, the regulation of PIAS protein activity in cells is poorly understood. Previously, we reported that expression of Trim32, a RING domain ubiquitin-protein isopeptide ligase-ubiquitin ligase mutated in human limb-girdle muscular dystrophy type 2H (LGMD2H) and Bardet-Biedl syndrome, is elevated during mouse skin carcinogenesis, protecting keratinocytes from apoptosis induced by UVB and tumor necrosis factor-α (TNFα). Here we report that Trim32 interacts with Piasy and promotes Piasy ubiquitination and degradation. Ubiquitination of Piasy by Trim32 could be reproduced in vitro using purified components. Their interaction was induced by treatment with UVB/TNFα and involved redistribution of Piasy from the nucleus to the cytoplasm, where it accumulated in cytoplasmic granules that colocalized with Trim32. Piasy destabilization and ubiquitination required an intact RING domain in Trim32. The LGMD2H-associated missense point mutation prevented Trim32 binding to Piasy, and human Piasy failed to colocalize with human Trim32 in fibroblasts isolated from an LGMD2H patient. Trim32 expression increased the transcriptional activity of NFκB in epidermal keratinocytes, both under basal treatment and after UVB/TNFα treatment. Conversely, Piasy inhibited NFκB activity under the same conditions and sensitized keratinocytes to apoptosis induced by TNFα and UVB. Our results indicate that, by controlling Piasy stability, Trim32 regulates UVB-induced keratinocyte apoptosis through induction of NFκB and suggests loss of function of Trim32 in LGMD2H.

Original languageEnglish (US)
Pages (from-to)25850-25866
Number of pages17
JournalJournal of Biological Chemistry
Volume281
Issue number35
DOIs
StatePublished - Sep 1 2006

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Ubiquitin-Protein Ligases
Keratinocytes
Protein Inhibitors of Activated STAT
Ubiquitination
Tumor Necrosis Factor-alpha
Apoptosis
Degradation
Ligases
Ubiquitin
Bardet-Biedl Syndrome
Cytoplasmic Granules
Missense Mutation
Fibroblasts
Point Mutation
Skin
Carcinogenesis
Cytoplasm
Transcription Factors
Therapeutics
Limb-girdle muscular dystrophy type 2H

ASJC Scopus subject areas

  • Biochemistry

Cite this

The interaction of piasy with Trim32, an E3-ubiquitin ligase mutated in limb-girdle muscular dystrophy type 2H, promotes piasy degradation and regulates UVB-induced keratinocyte apoptosis through NFκB. / Albor, Amador; El-Hizawi, Sally; Horn, Elizabeth J.; Laederich, Melanie; Frosk, Patrick; Wrogemann, Klaus; Kulesz-Martin, Molly.

In: Journal of Biological Chemistry, Vol. 281, No. 35, 01.09.2006, p. 25850-25866.

Research output: Contribution to journalArticle

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abstract = "Protein inhibitors of activated STATs (PIAS) family members are ubiquitin-protein isopeptide ligase-small ubiquitin-like modifier ligases for diverse transcription factors. However, the regulation of PIAS protein activity in cells is poorly understood. Previously, we reported that expression of Trim32, a RING domain ubiquitin-protein isopeptide ligase-ubiquitin ligase mutated in human limb-girdle muscular dystrophy type 2H (LGMD2H) and Bardet-Biedl syndrome, is elevated during mouse skin carcinogenesis, protecting keratinocytes from apoptosis induced by UVB and tumor necrosis factor-α (TNFα). Here we report that Trim32 interacts with Piasy and promotes Piasy ubiquitination and degradation. Ubiquitination of Piasy by Trim32 could be reproduced in vitro using purified components. Their interaction was induced by treatment with UVB/TNFα and involved redistribution of Piasy from the nucleus to the cytoplasm, where it accumulated in cytoplasmic granules that colocalized with Trim32. Piasy destabilization and ubiquitination required an intact RING domain in Trim32. The LGMD2H-associated missense point mutation prevented Trim32 binding to Piasy, and human Piasy failed to colocalize with human Trim32 in fibroblasts isolated from an LGMD2H patient. Trim32 expression increased the transcriptional activity of NFκB in epidermal keratinocytes, both under basal treatment and after UVB/TNFα treatment. Conversely, Piasy inhibited NFκB activity under the same conditions and sensitized keratinocytes to apoptosis induced by TNFα and UVB. Our results indicate that, by controlling Piasy stability, Trim32 regulates UVB-induced keratinocyte apoptosis through induction of NFκB and suggests loss of function of Trim32 in LGMD2H.",
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T1 - The interaction of piasy with Trim32, an E3-ubiquitin ligase mutated in limb-girdle muscular dystrophy type 2H, promotes piasy degradation and regulates UVB-induced keratinocyte apoptosis through NFκB

AU - Albor, Amador

AU - El-Hizawi, Sally

AU - Horn, Elizabeth J.

AU - Laederich, Melanie

AU - Frosk, Patrick

AU - Wrogemann, Klaus

AU - Kulesz-Martin, Molly

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AB - Protein inhibitors of activated STATs (PIAS) family members are ubiquitin-protein isopeptide ligase-small ubiquitin-like modifier ligases for diverse transcription factors. However, the regulation of PIAS protein activity in cells is poorly understood. Previously, we reported that expression of Trim32, a RING domain ubiquitin-protein isopeptide ligase-ubiquitin ligase mutated in human limb-girdle muscular dystrophy type 2H (LGMD2H) and Bardet-Biedl syndrome, is elevated during mouse skin carcinogenesis, protecting keratinocytes from apoptosis induced by UVB and tumor necrosis factor-α (TNFα). Here we report that Trim32 interacts with Piasy and promotes Piasy ubiquitination and degradation. Ubiquitination of Piasy by Trim32 could be reproduced in vitro using purified components. Their interaction was induced by treatment with UVB/TNFα and involved redistribution of Piasy from the nucleus to the cytoplasm, where it accumulated in cytoplasmic granules that colocalized with Trim32. Piasy destabilization and ubiquitination required an intact RING domain in Trim32. The LGMD2H-associated missense point mutation prevented Trim32 binding to Piasy, and human Piasy failed to colocalize with human Trim32 in fibroblasts isolated from an LGMD2H patient. Trim32 expression increased the transcriptional activity of NFκB in epidermal keratinocytes, both under basal treatment and after UVB/TNFα treatment. Conversely, Piasy inhibited NFκB activity under the same conditions and sensitized keratinocytes to apoptosis induced by TNFα and UVB. Our results indicate that, by controlling Piasy stability, Trim32 regulates UVB-induced keratinocyte apoptosis through induction of NFκB and suggests loss of function of Trim32 in LGMD2H.

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