Characterization of C. elegans RING finger protein 1, a binding partner of ubiquitin-conjugating enzyme 1

• Published in: Developmental biology Vol. 265; no. 2; pp. 446 - 459
• Main Authors: Crowe, Emily, Candido, E.Peter M
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 15-01-2004
• Subjects: Amino Acid Sequence; Animals; C. elegans; Caenorhabditis elegans; Caenorhabditis elegans - genetics; Caenorhabditis elegans - metabolism; Caenorhabditis elegans Proteins - genetics; Caenorhabditis elegans Proteins - metabolism; Carrier Proteins - genetics; Carrier Proteins - metabolism; Molecular Sequence Data; Precipitin Tests; Protein Interaction Mapping; RAD6; RING; RNA Interference; ubc-1; ubiquitin-conjugating enzyme 1; Ubiquitin-Conjugating Enzymes - genetics; Ubiquitin-Conjugating Enzymes - metabolism; Ubiquitylation; Vulva; C. elegans; RNA interference; ubc-1; Ubiquitylation; RING; RAD6; Vulva; E2
• ISSN: 0012-1606; 1095-564X
• DOI: 10.1016/j.ydbio.2003.09.037

In a yeast two-hybrid screen, RING finger protein 1 (RFP-1) and UBR1 were identified as potential binding partners of
C. elegans UBC-1, a ubiquitin-conjugating enzyme with a high degree of identity to
S. cerevisiae UBC2/RAD6. The interaction of RFP-1 and UBC-1 was confirmed by co-immunoprecipitation experiments. Yeast interaction trap experiments mapped the region of interaction to the basic N-terminal 313 residues of RFP-1. The acidic carboxy-terminal extension of UBC-1 was not required for the interaction with RFP-1. Western blot analysis and indirect immunohistochemical staining show that RFP-1 is present in embryos, larvae, and adults, where it is found in intestinal, nerve ring, pharyngeal, gonadal, and oocyte cell nuclei. Double-stranded RNA interference experiments against
rfp-1 indicate that this gene is required for L1 development, vulval development, and for egg laying. By contrast, RNA interference against
ubc-1 gave no obvious phenotype, suggesting that
ubc-1 is nonessential or is functionally redundant.