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Eukaryotic Cell, December 2004, p. 1433-1444, Vol. 3, No. 6
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.6.1433-1444.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Potential Link between the NIMA Mitotic Kinase and Nuclear Membrane Fission during Mitotic Exit in Aspergillus nidulans

Jonathan R. Davies,1 Aysha H. Osmani,1 Colin P. C. De Souza,1 Catherine Bachewich,2 and Stephen A. Osmani1*

Department of Molecular Genetics, The Ohio State University, Columbus, Ohio,1 Health Sector, Biotechnology Research Institute, National Research Council of Canada, Montreal, Quebec, Canada2

Received 21 July 2004/ Accepted 7 September 2004

We have isolated TINC as a NIMA-interacting protein by using the yeast two-hybrid system and have confirmed that TINC interacts with NIMA in Aspergillus nidulans. The TINC-NIMA interaction is stabilized in the absence of phosphatase inhibitors and in the presence of kinase-inactive NIMA, suggesting that the interaction is enhanced when NIMA is not fully activated. TINC is a cytoplasmic protein. TINC homologues and a TINC-like protein (A. nidulans HETC) are conserved in other filamentous fungi. Neither deletion of tinC nor deletion of both tinC and A. nidulans hetC is lethal, but deletion of tinC does produce cold sensitivity as well as osmotic sensitivity. Expression of an amino-terminal-truncated form of TINC ({Delta}N-TINC) inhibits colony growth in Aspergillus and localizes to membrane-like structures within the cell. Examination of cell cycle progression in these cells reveals that they progress through multiple defective mitoses. Many cells contain large polyploid single nuclei, while some appear to have separated masses of DNA. Examination of the nuclear envelopes of cells containing more than one DNA mass reveals that both DNA masses are contained within a single nuclear envelope, indicating that nuclear membrane fission is defective. The ability of these cells to separate DNA segregation from nuclear membrane fission suggests that this coordination is normally a regulated process in A. nidulans. Additional experiments demonstrate that expression of {Delta}N-TINC results in premature NIMA disappearance in mitotic samples. We propose that TINC's interaction with NIMA and the cell cycle defects produced by {Delta}N-TINC expression suggest possible roles for TINC and NIMA during nuclear membrane fission.

* Corresponding author. Mailing address: Department of Molecular Genetics, The Ohio State University, 804 Riffe Building, 496 W. 12th Ave., Columbus, OH 43210. Phone: (614) 247-6791. Fax: (614) 247-6845. E-mail: osmani.2{at}osu.edu.

Eukaryotic Cell, December 2004, p. 1433-1444, Vol. 3, No. 6
1535-9778/04/$08.00+0     DOI: 10.1128/EC.3.6.1433-1444.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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