Competency for nonsense-mediated reduction in collagen X mRNA is specified by the 3' UTR and corresponds to the position of mutations in Schmid metaphyseal chondrodysplasia.

Nonsense-mediated decay (NMD) is a eukaryotic cellular RNA surveillance and quality-control mechanism that degrades mRNA containing premature stop codons (nonsense mutations) that otherwise may exert a deleterious effect by the production of dysfunctional truncated proteins. Collagen X (COL10A1) nonsense mutations in Schmid-type metaphyseal chondrodysplasia are localized in a region toward the 3' end of the last exon (exon 3) and result in mRNA decay, in contrast to most other genes in which terminal-exon nonsense mutations are resistant to NMD. We introduce nonsense mutations into the mouse Col10a1 gene and express these in a hypertrophic-chondrocyte cell line to explore the mechanism of last-exon mRNA decay of Col10a1 and demonstrate that mRNA decay is spatially restricted to mutations occurring in a 3' region of the exon 3 coding sequence; this region corresponds to where human mutations have been described. This localization of mRNA-decay competency suggested that a downstream region, such as the 3' UTR, may play a role in specifying decay of mutant Col10a1 mRNA containing nonsense mutations. We found that deleting any of the three conserved sequence regions within the 3' UTR (region I, 23 bp; region II, 170 bp; and region III, 76 bp) prevented mutant mRNA decay, but a smaller 13 bp deletion within region III was permissive for decay. These data suggest that the 3' UTR participates in collagen X last-exon mRNA decay and that overall 3' UTR configuration, rather than specific linear-sequence motifs, may be important in specifying decay of Col10a1 mRNA containing nonsense mutations.

Hepatic and extrahepatic expression of vitellogenin genes in the zebrafish, Danio rerio.

Vitellogenins (Vtgs) are yolk precursor proteins in oviparous species and are cleaved into three portions-lipovitellin I (LVI), phosvitin (PV), and lipovitellin II (LVII)-in oocytes in vertebrates. In the present study, we found that the zebrafish genome contains at least seven vtg genes (vtg1-7) encoding heterogeneous vitellogenins with three distinct types of Vtgs: type I (Vtg1, 4-7) contains all the three major portions but lacks the C-terminal half of LVII; type II (Vtg2) is the only one including intact three portions; type III (Vtg3) lacks both PV and the LVII C-terminal half. The seven vtgs were located in two different chromosomes: one (vtg3) in LG11 and the rest closely linked in LG22, probably arisen from local gene duplication events. All of the seven vtgs are predominantly expressed in female liver and can be induced in male liver by 17beta-estradiol (E2). The level of vtg1 mRNA was about 100x and 1000x higher than those of vtg2 and vtg3 mRNAs. We also found vtg mRNAs in several non-liver tissues, but the expression level is generally <10% of that in the liver. In situ hybridization experiments confirmed that the extrahepatic expression was actually in adipocytes associated with several organs such as the intestine, ovary, and E2-induced testis.

Expression and localization of the Parkin co-regulated gene in mouse CNS suggests a role in ependymal cilia function.

Parkin Co-Regulated Gene (PACRG) is a gene that shares a bi-directional promoter with the Parkinson's disease associated gene parkin. The functional role of PACRG is not well understood, although the gene has been associated with parkinsonian syndromes and more recently with eukaryotic cilia and flagella. We investigated the expression of Pacrg in the mouse brain by in situ hybridization and observed robust expression of Pacrg in the cells associated with the lateral, third and fourth ventricle, in addition to the aqueduct of Sylvius and choroid plexus. For all regions of Pacrg expression identified, strong expression was observed in the newborn period and this was maintained into adulthood. Immunohistochemical analysis showed that Pacrg was a component of the ependymal cells and cilia lining the ventricles. Based on our results and the previous association of PACRG homologues with cilia and flagella, we propose that Pacrg is a component of the ependymal cilia and may play an important role in motile cilia development and/or function in the CNS.