Abstract: In this paper, we describe RT-PCR strategies that allowed us to identify and clone members of the NEP-like family. Degenerate oligoncleotide primers corresponding to consensus sequences located on either side of the HEXXH consensus sequence for zincins were designed and used in RT-PCR with mouse and human testis cDNAs. DNA fragments with lengths expected from the sequence of this class of enzympes were obtained. These DNA fragments were cloned and sequenced. Using this PCR strategy and the PCR fragments as probes to screen cDNA libraries, three zincin-like peptidases were identified in addition of known members of the family. The cDNA sequences allowed to derive specific probes for Northern and in situ hybridization, and probe human chromosomes to localize the gene and establish potential links to genetic diseases. Furthermore, these cDNA sequences were used to produce recombinant fusion proteins in Escherichia coli in order to raise specific antibodies.

Abstract: The present invention relates to mammalian staufen, a double-stranded RNA-binding protein involved in mRNA transport and localization. The invention further relates to the demonstration of the association of a RNA-binding protein with the rough endoplasmic reticulum (RER), implicating staufen and related proteins in the transport of RNA to its site of translation. Broadly, the invention therefore relates to transport and translation of RNA. More specifically, the present invention relates to human and mouse staufen proteins and to the modulation of transport of RNA to the RER by these proteins. The present invention also relates to isolated nucleic acid molecules encoding mammalian staufen, as well as vectors and host cells harboring same. In addition, the present invention relates to screening assays for identifying modulators of staufen activity and to the identification of mutants thereof which abrogate their interaction with RER.

Abstract: The present invention relates to mammalian Staufen, a double-stranded RNA-binding protein involved in mRNA transport and localization. The invention further relates to the demonstration of the association of a RNA-binding protein with the rough endoplasmic reticulum (RER), implicating Staufen and related proteins in the transport of RNA to its site of translation. Broadly, the invention therefore relates to transport and translation of RNA. More specifically, the present invention relates to human and mouse Staufen proteins and to the modulation of transport of RNA to the RER by these proteins. The present invention also relates to isolated nucleic acid molecules encoding mammalian Staufen, as well as vectors and host cells harboring same. In addition, the present invention relates to screening assays for identifying modulators of Staufen activity and to the identification of mutants thereof which abrogate their interaction with RER.