Conserved Asf1-importin β physical interaction in growth and sexual development in the ciliate Tetrahymena thermophila.

Abstract

UNLABELLED How the eukaryotic cell specifies distinct chromatin domains is a central problem in molecular biology. The ciliate protozoan Tetrahymena thermophila features a separation of structurally and functionally distinct germ-line and somatic chromatin into two distinct nuclei, the micronucleus (MIC) and macronucleus (MAC) respectively. To address questions about how distinct chromatin states are assembled in the MAC and MIC, we have initiated studies to define protein-protein interactions for T. thermophila chromatin-related proteins. Affinity purification followed by mass spectrometry analysis of the conserved Asf1 histone chaperone in T. thermophila revealed that it forms a complex with an importin β, ImpB6. Furthermore, these proteins co-localized to both the MAC and MIC in growth and development. We suggest that newly synthesized histones H3 and H4 in T. thermophila are transported via Asf1-ImpB6 in an evolutionarily conserved pathway to both nuclei where they then enter nucleus-specific chromatin assembly pathways. These studies set the stage for further use of functional proteomics to elucidate details of the characterization and functional analysis of the unique chromatin domains in T. thermophila. BIOLOGICAL SIGNIFICANCE Asf1 is an evolutionarily conserved chaperone of H3 and H4 histones that functions in replication dependent and independent chromatin assembly. Although Asf1 has been well studied in humans and yeast (members of the Opisthokonta lineage of eukaryotes), questions remain concerning its mechanism of function. To obtain additional insight into the Asf1 function we have initiated a proteomic analysis in the ciliate protozoan T. thermophila, a member of the Alveolata lineage of eukaryotes. Our results suggest that an evolutionarily conserved function of Asf1 is mediating the nuclear transport of newly synthesized histones H3 and H4.

DOI: 10.1016/j.jprot.2013.09.018

Cite this paper

@article{Garg2013ConservedA, title={Conserved Asf1-importin β physical interaction in growth and sexual development in the ciliate Tetrahymena thermophila.}, author={Jyoti Garg and Jean-Philippe Lambert and Abdel R Karsou and Susanna Marquez and Syed Nabeel-Shah and Virginia Bertucci and Dashaini V Retnasothie and Ernest Radovani and Tony Pawson and Anne-Claude Gingras and Ronald E . Pearlman and Jeffrey S. Fillingham}, journal={Journal of proteomics}, year={2013}, volume={94}, pages={311-26} }