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From Midbody Protein−Protein Interaction Network Construction to Novel Regulators in Cytokinesis

Supporting Info

Tzu-Chi Chen^†^#, Sheng-An Lee^§^#, Tse-Ming Hong ^#, Jin-Yuan Shih ^#, Jin-Mei Lai, Hsin-Ying Chiou, Shuenn-Chen Yang, Chen-Hsiung Chan^§, Cheng-Yan Kao^§, Pan-Chyr Yang* and Chi-Ying F. Huang*^†^‡^§

Institute of Biotechnology in Medicine, Institute of Clinical Medicine, National Yang-Ming University, Taipei 112, Taiwan, R.O.C., Department of Computer Science and Information Engineering, National Taiwan University, Taipei 106, Taiwan, R.O.C., NTU Center for Genomic Medicine, College of Medicine, National Taiwan University, Taipei 100, Taiwan, R.O.C., Department of Internal Medicine, National Taiwan University Hospital, Taipei 100, Taiwan, R.O.C., Department of Life Science, Fu-Jen Catholic University, Taipei Hsien 242, Taiwan, R.O.C., and Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, R.O.C.

J. Proteome Res., 2009, 8 (11), pp 4943–4953

DOI: 10.1021/pr900325f

Publication Date (Web): October 2, 2009

* Additional corresponding author: Pan-Chyr Yang. E-mail: pcyang@ntu.edu.tw; Contact Information: Correspondence: Chi-Ying F. Huang, Professor, Institute of Clinical Medicine, National Yang-Ming University, Taipei 112, Taiwan, Phone: 886-228267904, Fax: 886-228745074, E-mail: cyhuang5@ym.edu.tw., †

Institute of Biotechnology in Medicine, National Yang-Ming University.

, #

These authors contributed equally to this work.

, §

Department of Computer Science and Information Engineering, National Taiwan University.

NTU Center for Genomic Medicine, College of Medicine, National Taiwan University.

Department of Internal Medicine, National Taiwan University Hospital.

Fu-Jen Catholic University.

Institute of Biomedical Sciences, Academia Sinica.

, ‡

Institute of Clinical Medicine, National Yang-Ming University.

Abstract

Midbody, a transient organelle-like structure, is known as central for abscission and is indispensable for termination of cytokinesis. Here, we used the midbody proteome inventories to construct the potential midbody protein−protein interaction (PPI) network. To delineate novel regulators participating in cytokinesis, the z-score, a standard statistic score, rather than hub degree was implemented to prioritize the novel hubs. Of these hubs, KIAA0133, SEPT1, KIAA1377, and CRMP-1 were localized to the midbody, whereas HTR3A and ICAM2 were associated with the cleavage furrow as examined by immunofluorescence. Knockdown of SEPT1 and KIAA1377 resulted in increasing numbers of cytokinesis defect cells, suggesting these newly identified hubs play critical roles in cytokinesis progression. Moreover, ectopic expression of CRMP-1 mutant in which Aurora-A phosphorylation sites have been replaced with Ala results in a cytokinesis defect. This subproteome network construction not only sheds light on the intimate interactions of the midbody proteomes, but also prioritizes novel hubs or protein complexes that may govern the process of cytokinesis.

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