Mutations affecting spindle pole body and mitotic exit network function are synthetically lethal with a deletion of the nucleoporin NUP1 in S. cerevisiae.
Nuclear pore complexes (NPCs) are embedded in the nuclear envelope of eukaryotic cells and function to regulate passage of macromolecules in and out of the nucleus. Nup1 is one of 30 nucleoporins comprising the NPC of the yeast Saccharomyces cerevisiae and is located on the nucleoplasmic face of the NPC where it plays a role in mRNA export and protein transport. In order to further characterize the function of Nup1 we used a genetic approach to identify mutations that are synthetically lethal in combination with a deletion of NUP1 (nup1Delta). We have identified one such nup1 lethal mutant (nle6) as a temperature sensitive allele of nud1. NUD1 encodes a component of the yeast spindle pole body (SPB) and acts as scaffolding for the mitotic exit network (MEN). We observe that nle6/nud1 mutant cells have a normal distribution of NPCs within the nuclear envelope and exhibit normal rates of nuclear protein import at both the permissive and restrictive temperatures. nup1Delta also exhibits synthetic lethality with bub2Delta and bfa1Delta, both of which encode proteins that colocalize with Nud1 at spindle pole bodies and function in the mitotic exit network. However, we do not observe genetic interactions among nle6/nud1, bub2Delta, or bfa1Delta and mutations in the nucleoporin encoding genes NUP60 or NUP170, nor is nup1Delta synthetically lethal with the absence of components downstream in the mitotic exit network, including Lte1, Swi5, and Dbf2. Our results suggest a novel functional connection between Nup1 and proteins comprising both the spindle pole body and early mitotic exit network.
Harper
Nicola C
NC
Department of Biology, Colgate University, NY 13346, USA.
Al-Greene
Nicole T
NT
Basrai
Munira A
MA
Belanger
Kenneth D
KD
D003851:Deoxyribonucleases / Q000235:genetics; D017353:Gene Deletion; D009154:Mutation; D022022:Nuclear Pore / Q000737:chemistry; D028861:Nuclear Pore Complex Proteins / Q000172:deficiency / Q000235:genetics*; D009687:Nuclear Proteins / Q000235:genetics; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics; D008941:Spindle Apparatus / Q000737:chemistry / Q000235:genetics*; D012359:tRNA Methyltransferases
C064550:NUP1 protein, S cerevisiae; D028861:Nuclear Pore Complex Proteins; D009687:Nuclear Proteins; D029701:Saccharomyces cerevisiae Proteins; D012359:tRNA Methyltransferases; D003851:Deoxyribonucleases; C077501:TRM2 protein, S cerevisiae