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Mol Cell Biol, March 1998, p. 1303-1311, Vol. 18, No. 3
0270-7306/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

The Alpha Chain of the Nascent Polypeptide-Associated Complex Functions as a Transcriptional Coactivator

Wagner V. Yotov,dagger Alain Moreau,Dagger and René St-Arnaud*

Genetics Unit, Shriners Hospital, and Departments of Surgery and Human Genetics, McGill University, Montréal, Québec H3G 1A6, Canada

Received 10 June 1997/Returned for modification 15 August 1997/Accepted 24 November 1997

We report the characterization of clone 1.9.2, a gene expressed in mineralizing osteoblasts. Remarkably, clone 1.9.2 is the murine homolog of the alpha chain of the nascent polypeptide-associated complex (alpha -NAC). Based on sequence similarities between alpha -NAC/1.9.2 and transcriptional regulatory proteins and the fact that the heterodimerization partner of alpha -NAC was identified as the transcription factor BTF3b (B. Wiedmann, H. Sakai, T. A. Davis, and M. Wiedmann, Nature 370:434-440, 1994), we investigated a putative role for alpha -NAC/1.9.2 in transcriptional control. The alpha -NAC/1.9.2 protein potentiated by 10-fold the activity of the chimeric activator GAL4/VP-16 in vivo. The potentiation was shown to be mediated at the level of gene transcription, because alpha -NAC/1.9.2 increased GAL4/VP-16-mediated mRNA synthesis without affecting the half-life of the GAL4/VP-16 fusion protein. Moreover, the interaction of alpha -NAC/1.9.2 with a transcriptionally defective mutant of GAL4/VP-16 was severely compromised. Specific protein-protein interactions between alpha -NAC/1.9.2 and GAL4/VP-16 were demonstrated by gel retardation, affinity chromatography, and protein blotting assays, while interactions with TATA box-binding protein (TBP) were detected by immunoprecipitation, affinity chromatography, and protein blotting assays. Based on these interactions that define the coactivator class of proteins, we conclude that the alpha -NAC/1.9.2 gene product functions as a transcriptional coactivator.

* Corresponding author. Mailing address: Genetics Unit, Shriners Hospital, 1529 Cedar Ave., Montréal, Québec H3G 1A6, Canada. Phone: (514) 842-5964. Fax: (514) 842-5581. E-mail: rst-arnaud{at}shriners.mcgill.ca.

dagger Present address: Centre de cancérologie Charles Bruneau, Hôpital Ste-Justine, Montréal, Québec H3T 1C5, Canada.

Dagger Present address: Laboratoire de génétique moléculaire, Institut de Recherches Cliniques de Montréal, Montréal, Québec H2W 1R7, Canada.




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