The most significant result of IMPG SAS of 2018 in basic research

The scientific board of CBs SAS evaluated as the 1st most important result of 2018 in the basic research the work of the authors J. Kralovicova and I. Sevcikova:

‘PUF60-activated exons uncover altered 3 ‘; splice-site selection by germline missense mutations in a single RRM’

Eukaryotic genes contain introns that must be removed from the precursor mediator RNA using a large and highly dynamic spliceosome complex. The spliceosome is formed on each intron, including U1, U2 and U4 / 5/6 small nuclear ribonucleoproteins and a number of other proteins. The critical step in the formation of the splicing apparatus is the interaction of U1 with the 5 ′ splicing apparatus and a U2 with a branch point assisted by a U2 helper factor (U2AF) binding to a 3 ′ splice site (3′ss). PUF60 is one of the splicing facts that are homologous to U2AF. The PUF60 molecule contains two RNA recognition motifs (RRM) and facilitates the association of U2 with the primary transcript in cooperation with U2AF. The deficiency of PUF60 (PD) is associated with delayed development, mental disability and renal, spinal cord and cardiac disorders, but the pathogenesis of the disease has not yet been elucidated. Using RNA NGS, we have identified and characterized genes / exons regulated by the PUF60 protein, which acts predominantly as their activator. Using co-transfection of splice minigens with PUF60 constructs, we observed the functional consequences of PDF60 mutations associated with PD. We have found that mutations affect the selection of competing 3 PU splice sites regulated by PUF60, and that the heterogeneity of germline mutations located in the same PUF60 RRM may increase phenotypic variability at the splice site selection level. Project: VEGA 2/0057/18 Publication: KRALOVICOVA, JanaSEVCIKOVA, Ivana – STEJSKALOVA, Eva – OBUCA, Mina – HILLER, Michael – STANEK, David – VORECHOVSKY, Igor. PUF60-activated exons uncover altered 3 ‘ splice-site selection by germline missense mutations in a single RRM. In Nucleic acids research, 2018, vol. 46, no. 12, p. 6166-6187. (11.561 – IF2017). ISSN 0305-1048.