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Figure 5


Fig. 5. Characterization of the CrLIS1 gene from Chlamydomonas. (A) Southern blot analysis of Chlamydomonas genomic DNA using a CrLIS1-specific probe. The blot demonstrates that CrLIS1 is present in a single copy in the Chlamydomonas genome. (B) Northern blot analysis of Chlamydomonas mRNA showing upregulation of the ~2.2 kb CrLIS1 mRNA in deflagellated cells (30'postDF) relative to non-deflagellated cells (NDF). (C) SMART analysis (http://smart.emblheidelberg.de/) of the human Lis1 and CrLis1 polypeptide sequences (see Fig. 4) showing that CrLis1 contains seven WD repeats, as does the human Lis1 protein. However, note the absence of the N-terminal ~30-residue LisH domain in CrLis1. (D) Phylogenetic analysis of CrLis1 based on the ClustalW alignment of WD-repeat proteins (Fig. 4); proteins included in the analysis are Chlamydomonas ODA IC1 and IC2, rat cytoplasmic dynein IC74, human Gbeta 1, human Lis1 and Chlamydomonas CrLis1. CrLis1 is most closely related to mammalian Lis1.