Identification and characterisation of structural maintenance of chromosome 1 (smc1) mutants of Coprinopsis cinerea.
A Coprinopsis cinerea homokaryotic fruiting strain was mutagenised, identifying a mutant that exhibited a hyphal growth temperature sensitive defect and hyphal knot development defect at an early fruiting stage, even at the hyphal growth permissive temperature. Microscopic observation suggested that the mutant nuclei exhibited defects in the metaphase to anaphase transition at the restrictive temperature. The gene in which the mutation occurred was cloned, sequenced and determined to be homologous to smc1. Sequence analyses of the mutant revealed deletion of 28 base pairs in the 19th intron of the Cc.smc1 gene, resulting in complete failure of splicing of that intron and in insertion of 14 amino acids in the C-terminal region of the Cc.Smc1 protein. We isolated eight hyphal growth revertants and identified four intragenic suppressors. All were the result of amino acid substitutions in the C-terminal region. Three of the suppressors caused reversion of the arrest in an early fruiting stage. One of the suppressors exhibited cold sensitivity and failed to suppress the fruiting defect, suggesting that flexibility of a lobe in the C-terminal region is important for proper function of Cc.Smc1.
Muraguchi
Hajime
H
Department of Biotechnology, Faculty of Bioresource Sciences, Akita Prefectural University, Shimoshinjo-nakano, Akita, Japan. muraguchi@akita-pu.ac.jp
Abe
Kousuke
K
Nakagawa
Minako
M
Nakamura
Kiyoto
K
Yanagi
Sonoe O
SO
D000595:Amino Acid Sequence; D002453:Cell Cycle; D018797:Cell Cycle Proteins / Q000737:chemistry / Q000235:genetics*; D002868:Chromosomal Proteins, Non-Histone / Q000737:chemistry / Q000235:genetics*; D003302:Coprinus / Q000166:cytology / Q000235:genetics*; D005800:Genes, Fungal; D025301:Hyphae / Q000166:cytology / Q000254:growth & development; D008969:Molecular Sequence Data; D050505:Mutant Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009154:Mutation / Q000235:genetics*; D017433:Protein Structure, Secondary; D013172:Spores, Fungal / Q000166:cytology / Q000502:physiology; D013489:Suppression, Genetic; D013696:Temperature; D000098550:Structural Maintenance of Chromosome Protein 1
D018797:Cell Cycle Proteins; D002868:Chromosomal Proteins, Non-Histone; D050505:Mutant Proteins; D000098550:Structural Maintenance of Chromosome Protein 1