Aspergillus nidulans class V and VI chitin synthases CsmA and CsmB, each with a myosin motor-like domain, perform compensatory functions that are essential for hyphal tip growth.
The polarized synthesis of cell wall components such as chitin is essential for the hyphal tip growth of filamentous fungi. The actin cytoskeleton is known to play important roles in the determination of hyphal polarity in Aspergillus nidulans. Previously, we suggested that CsmA, a chitin synthase with a myosin motor-like domain (MMD), was involved in polarized chitin synthesis in a manner dependent on the interaction between the MMD and the actin cytoskeleton. The genome database indicates that A. nidulans possesses another gene encoding another chitin synthase with an MMD. In this study, we characterized this gene, which we designated csmB. The csmB null mutants examined were viable, although they exhibited defective phenotypes, including the formation of balloons and intrahyphal hyphae and the lysis of subapical regions, which were similar to those obtained with csmA null mutants. Moreover, csmA csmB double null mutants were not viable. Mutants in which csmB was deleted and the expression of csmA was under the control of the alcA promoter were viable but severely impaired in terms of hyphal growth under alcA-repressing conditions. We revealed that CsmB with three copies of a FLAG epitope tag localized at the hyphal tips and forming septa, and that the MMD of CsmB was able to bind to actin filaments in vitro. These results suggest that CsmA and CsmB perform compensatory functions that are essential for hyphal tip growth.
Department of Biotechnology, The University of Tokyo, 1-1-1 Yayoi, Tokyo 113-8657, Japan.
Norio
N
Takeshita
Shuichi
S
Yamashita
Akinori
A
Ohta
Hiroyuki
H
Horiuchi
D000199:Actins / Q000378:metabolism; D001233:Aspergillus nidulans / Q000201:enzymology* / Q000378:metabolism / Q000502:physiology; D049109:Cell Proliferation; D002687:Chitin Synthase / Q000235:genetics / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D005656:Fungal Proteins / Q000235:genetics / Q000378:metabolism; D015966:Gene Expression Regulation, Fungal; D020409:Molecular Motor Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009154:Mutation; D009218:Myosins / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D014158:Transcription, Genetic
D000199:Actins; D005656:Fungal Proteins; D020409:Molecular Motor Proteins; D002687:Chitin Synthase; D009218:Myosins