Domain definition and interaction mapping for the endonuclease complex hNob1/hPno1

ABSTRACT

Ribosome biogenesis requires a variety of trans-acting factors in order to produce functional ribosomal subunits. In human cells, the complex formed by the proteins hNob1 and hPno1 is crucial to the site 3 cleavage occurring at the 3ʹ-end of 18S pre-rRNA. However, the properties and activity of this complex are still poorly understood. We present here a detailed characterization of hNob1 organization and its interaction with hPno1. We redefine the boundaries of the endonuclease PIN domain present in hNob1 and we further delineate the precise interacting modules required for complex formation in hNob1 and hPno1. Altogether, our data contributes to a better understanding of the complex biology required during the site 3 cleavage step in ribosome biogenesis.

KEYWORDS:

• Ribosome biogenesis
• endonuclease
• pre-rRNA
• KH domain
• PIN domain
• zinc ribbon

Acknowledgments

We thank Cameron Mackereth and Fabien Darfeuille for critical reading of the manuscript. We further acknowledge Simon Lebaron, Marie-Françoise O’Donohue, Lucile Maleuvre and Manon Coursières for experiments, which unfortunately could not be included. This work was funded by ANR grant (ANR Blanc 2010 grant RIBOPRE40S (ANR-10-BLAN-1224) and RIBOMAN ANR-16-CE11-0029 to S.F.), the INSERM and the University of Bordeaux.

Disclosure statement

No potential conflict of interest was reported by the authors.

Additional information

Funding

This work was supported by the Agence Nationale de la Recherche [ANR-16-CE11-0029]; Agence Nationale de la Recherche [ANR-10-BLAN-1224].