Characterisation of the nascent polypeptide-associated complex in fission yeast.
The nascent polypeptide-associated complex (NAC) is an abundant and phylogenetically conserved protein complex. It is composed of two subunits and interacts with nascent polypeptide chains emerging from the ribosome. It has been proposed to protect the nascent chains from premature interaction with other cell proteins, but has also been found to associate with DNA junctions, and to be involved in other processes including transcription regulation and mitochondrial protein import.Here, we characterize NAC in fission yeast. We find that NAC is associated with ribosomes, while a significant fraction remains in a free form. The NAC alpha subunit contains a ubiquitin-associated (UBA) domain, which is found in several proteins involved in the ubiquitin-proteasome pathway for protein degradation. However, NAC does not associate with ubiquitin chains and mutants lacking NAC did not exhibit any obvious defects in protein degradation. Accordingly, we find that the NAC UBA domain belongs to an ancient and distinct subgroup of the UBA family. In contrast to the situation with budding yeast, fission yeast cells devoid of NAC were not temperature sensitive. However, they displayed resistance to the amino acid analogue canavanine, in accordance with the idea that NAC is involved in protein quality control.
Andersen
Katrine M
KM
Institute for Molecular Biology and Physiology, University of Copenhagen, Universitetsparken 13, 2100 Copenhagen Ø, Denmark.
Semple
Colin A
CA
Hartmann-Petersen
Rasmus
R
D000595:Amino Acid Sequence; D002172:Canavanine / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects; D025141:Drug Resistance, Fungal / Q000235:genetics; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism*; D018832:Molecular Chaperones / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009154:Mutation; D010802:Phylogeny; D011132:Polyribosomes / Q000378:metabolism; D011485:Protein Binding / Q000187:drug effects; D012270:Ribosomes / Q000378:metabolism; D012568:Schizosaccharomyces / Q000145:classification / Q000235:genetics / Q000378:metabolism*; D017386:Sequence Homology, Amino Acid; D025801:Ubiquitin / Q000378:metabolism
D005656:Fungal Proteins; D018832:Molecular Chaperones; D025801:Ubiquitin; C096099:nascent-polypeptide-associated complex; D002172:Canavanine