Small ubiquitin-like modifier-2 modification of retinoic acid receptor-alpha regulates its subcellular localization and transcriptional activity

Li Zhu; Nadine C Santos; Kwan Hee Kim

doi:10.1210/en.2009-0868

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Small ubiquitin-like modifier-2 modification of retinoic acid receptor-alpha regulates its subcellular localization and transcriptional activity

Journal article

Open access

Peer reviewed

Small ubiquitin-like modifier-2 modification of retinoic acid receptor-alpha regulates its subcellular localization and transcriptional activity

Li Zhu, Nadine C Santos and Kwan Hee Kim

Endocrinology (Philadelphia), Vol.150(12), pp.5586-5595

12/2009

DOI: https://doi.org/10.1210/en.2009-0868

Handle:

https://hdl.handle.net/2376/108457

PMCID: PMC2795702

PMID: 19850744

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Abstract

Transcriptional Activation - genetics

Humans

Transcriptional Activation - drug effects

Cercopithecus aethiops

Molecular Sequence Data

Male

Receptors, Retinoic Acid - genetics

Small Ubiquitin-Related Modifier Proteins - genetics

Cell Nucleus - metabolism

Sertoli Cells - cytology

Lysine - metabolism

Tretinoin - pharmacology

Sertoli Cells - metabolism

Amino Acid Sequence

Cell Line

Small Ubiquitin-Related Modifier Proteins - metabolism

Rats

Receptors, Retinoic Acid - metabolism

Binding Sites - genetics

Reverse Transcriptase Polymerase Chain Reaction

Rats, Sprague-Dawley

Blotting, Western

Sequence Homology, Amino Acid

Lysine - genetics

Animals

Protein Binding

Protein Processing, Post-Translational

Mutation

Cell Nucleus - drug effects

COS Cells

Retinoic Acid Receptor alpha

The retinoic acid receptor-alpha (Rara) gene is critical for germ cell development in the testis, as demonstrated by infertile Rara knockout male mice. The encoded protein for Rara (RARA) is expressed in both Sertoli cells and germ cells, but it is not always in the nucleus. Previously, all-trans retinoic acid (ATRA) was shown to increase the nuclear localization and transcriptional activity of RARA in Sertoli cells. Here, we identified a small ubiquitin-like modifier-2 (SUMO-2) modification as a novel posttranslational regulatory mechanism controlling the ATRA-dependent RARA subcellular localization and transcription. ATRA increased the SUMO-2 modification of RARA. In the presence of ATRA, lysine 166 (K166) and K171 of RARA were modified at a physiological concentration of SUMO-2, whereas in the absence of ATRA, K399 was the only site that was modified, but at a higher SUMO-2 concentration. However, K399 was critical for ATRA-controlled nuclear trafficking of RARA. In the presence of ATRA, a K399 mutation to arginine resulted in the cytoplasmic localization of K399R mutant, indicating that K166 and K171 sumoylations were inhibitory to nuclear localization. This may be due to SUMO/sentrin-specific peptidase 6 (SENP6) not being able to bind K399R mutant to desumoylate K166 and K171 in Sertoli cells, whereas it can bind RARA with intact K399. On the other hand, functional K166 and K171 sites for sumoylation were required for a full transcriptional activity, when K399 was intact. These results together suggest that both K166 and K171 sumoylation and desumoylation are critical for optimal RARA function.

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Details

Title: Small ubiquitin-like modifier-2 modification of retinoic acid receptor-alpha regulates its subcellular localization and transcriptional activity
Creators: Li Zhu - School of Molecular Biosciences, Center for Reproductive Biology, Washington State University, Pullman, Washington 99164, USA
Nadine C Santos
Kwan Hee Kim
Publication Details: Endocrinology (Philadelphia), Vol.150(12), pp.5586-5595
Publisher: United States
Grant note: R01 HD044569 / NICHD NIH HHS HD44569 / NICHD NIH HHS
Identifiers: 99900547523501842
Language: English
Resource Type: Journal article