Functional characterization of the microtubule-binding and -destabilizing domains of CPAP and d-SAS-4

Wen Bin Hsu, Liang-Yi Hung, Chieh Ju C. Tang, Chia Li Su, Yulin Chang, Tang K. Tang

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

We previously identified a novel centrosomal protein CPAP, which carries a 112-residue motif that is essential for microtubule destabilization. In this report, we define both the microtubule (MT) binding and destabilizing domains in human CPAP and analyze the mutations that affect its MT-destabilizing activity. Analysis of a series of CPAP truncated proteins showed that the MT-binding domain (MBD; residues 423-607) of CPAP is located next to its MT-destabilizing domain (MDD; residues 311-422). Site-specific mutagenesis revealed that the mutations that either disrupt the α-helical structure (Y341P, I346P, L348P, and triple-P) or alter the charge property (KR377EE) of the MDD significantly affect its MT-destabilizing ability. The activity for binding to a tubulin heterodimer was also significantly reduced in KR377EE mutant. Furthermore, we have analyzed the putative function of Drosophila d-SAS-4, a distant relative of human CPAP, which shares a conserved ∼ 20-aa sequence with the MDD of CPAP. Our results show that mutations in this conserved sequence also eliminate d-SAS-4′s MT-destabilizing activity, suggesting that d-SAS-4 and CPAP may play similar roles within cells.

Original languageEnglish
Pages (from-to)2591-2602
Number of pages12
JournalExperimental Cell Research
Volume314
Issue number14
DOIs
Publication statusPublished - 2008 Aug 15

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Microtubules
Mutation
Microtubule Proteins
Conserved Sequence
Tubulin
Site-Directed Mutagenesis
Drosophila
Proteins

All Science Journal Classification (ASJC) codes

  • Cell Biology

Cite this

Hsu, Wen Bin ; Hung, Liang-Yi ; Tang, Chieh Ju C. ; Su, Chia Li ; Chang, Yulin ; Tang, Tang K. / Functional characterization of the microtubule-binding and -destabilizing domains of CPAP and d-SAS-4. In: Experimental Cell Research. 2008 ; Vol. 314, No. 14. pp. 2591-2602.
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Functional characterization of the microtubule-binding and -destabilizing domains of CPAP and d-SAS-4. / Hsu, Wen Bin; Hung, Liang-Yi; Tang, Chieh Ju C.; Su, Chia Li; Chang, Yulin; Tang, Tang K.

In: Experimental Cell Research, Vol. 314, No. 14, 15.08.2008, p. 2591-2602.

Research output: Contribution to journalArticle

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AU - Hung, Liang-Yi

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