ENP1, an essential gene encoding a nuclear protein that is highly conserved from yeast to humans

A novel gene in Saccharomyces cerevisiae, ENP1, was found to be essential for growth. The ENP1 gene encodes a protein of 483 amino acids (aa). Nucleotide sequence analysis revealed that the deduced aa sequence of this gene exhibited approx. 60% sequence similarity to the deduced aa sequence of proteins of unknown function in Drosophila, Caenorhabditis elegans and humans. No well defined functional motifs were evident upon analysis of the aa sequence. The protein was found to contain 20% acidic aa residues, with most of them being localized to a very negatively charged domain between aa residues 100 and 150. A construct encoding a fusion protein consisting of the Enp1 protein fused to the c-myc epitope that was either under the control of the ENP1 promoter or the GAL1,10 promoter was prepared. The construct was used to express the protein tagged with the c-myc epitope. Despite the presence of a naturally occurring promoter region with homology to the unfolded protein response element, the level of Enp1mycp remained unchanged after growth of the cells in the presence of tunicamycin, an inhibitor of N-linked glycosylation of proteins. Immunohistochemical studies to define the cellular localization of the Enp1myc protein revealed that it was localized to the nucleus. Accession No.: U50779.