Characterization and in vivo functional analysis of the Schizosaccharomyces pombe ICln gene

ABSTRACT: During the early steps of snRNP biogenesis, the Survival of Motor Neuron (SMN) complex acts together with the methylosome, an entity formed by the pICln protein, WD45 and the PRMT5 methyltransferase. To expand our understanding of pICln and SMN functional relationships in vivo, we performed a genetic analysis of an uncharacterized S. pombe pICLn homologue. Although not essential, the S. pombe ICln protein is important for optimal yeast cell growth. The human pICln gene complements the iclnM-bM-^HM-^F slow growth phenotype demonstrating that the identified SpICln sequence represents the bona fide human homolog. Consistent with the role inferred for human pICln using in vitro experiments, we found that the SpICln protein is required for optimal production of the spliceosomal snRNPs and for efficient splicing in vivo. Genetic interaction approaches demonstrate furthermore that modulation of ICln activity is unable to compensate for defects induced by SMN mutations, and reciprocally. Using a genome-wide approach and RT-PCR validation tests, we show also that splicing is altered differentially in iclnM-bM-^HM-^F cells. Our data are consistent with the emerging view that splice site selection and spliceosome kinetics is highly dependent on the concentration of core spliceosomal components. RNA from M-NM-^TIcln mutant (2 replicates) vs RNA from wild type cells (2 replicates)

ORGANISM(S): Schizosaccharomyces pombe

SUBMITTER: johann Soret 

PROVIDER: E-GEOD-41403 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress