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Discovering the pre‐60S ribosome biogenesis factor interactome (560.7)
Author(s) -
McCann Kathleen,
Charette J. Michael,
Vincent Nicholas,
Baserga Susan
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.560.7
Subject(s) - ribosome biogenesis , interactome , ribosome , biology , ribosomal protein , eukaryotic ribosome , nucleolus , microbiology and biotechnology , ribosomal rna , helicase , computational biology , genetics , rna , cytoplasm , gene
Maturation of the 60S ribosomal subunit is a highly coordinated and dynamic process that begins in the nucleolus and ends in the cytoplasm. Over 80 different proteins, including several helicases, are found in the 60S pre‐ribosome and play a role in ribosome assembly. However, little is known about whether subcomplexes exist and how these subcomplexes function and regulate pre‐60S ribosome assembly. To begin to elucidate how the 60S pre‐ribosome is assembled, we performed three iterations of a high‐throughput yeast two‐hybrid analysis. We identified 234 high‐confidence interactions among 61 nucleolar proteins, representing a 7.5‐fold increase from current knowledge. Independent validation experiments confirm interaction in 98% of those tested. The 60s pre‐ribosome interactome has revealed several novel likely subcomplexes. One, formed from RNA helicases essential for growth, suggests the presence of a “helicasosome”. Glycerol gradient sedimentation analysis revealed that these helicases do, in fact, co‐peak as part of a 2 MDa complex. Furthermore, Dbp10, a component of the putative helicasosome, is found in smaller fractions on glycerol gradients suggesting that it is part of a smaller subcomplex as well. Since most of the yeast pre‐60S biogenesis factors are conserved to humans, many of the interactions and potential regulatory mechanisms that we will uncover are likely to be conserved as well. Grant Funding Source : NIHGM52581