Molecular cloning and characterization of a novel mouse actin-binding protein Zfp185
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Zinc finger protein (Zfp) 185 is a mouse protein containing a Lin-l1, Isl-1 and Mec-3 (LIM) domains at its C-terminus. It was recognized by comparing the genome sequence between humans and mice in 1997. In this study, we cloned the full-length Zfp185 by means of RACE and RT-PCR. Zfp185 may be closely associated with F-actin in cells as determined by a confocal microscopy. With a series of deletants of Zfp185 and GST-pull-down assay, we determined that N-terminus region (1–144) but not the LIM domain at C-terminus of Zfp185 protein was essential and sufficient to bind to F-actin cytoskeleton. Thus, our data offered evidence for the association of mouse Zfp185 with F-actin, which supports the potential role of Zfp185 in cell fundamental activity.
KeywordsZfp185 Clone Actin-binding
Zinc finger protein 185
Lin-l 1, Isl-1 and Mec-3
Rapid amplification of cDNA ends
Green fluorescent protein
We thank Dr. David E. Corbin for his careful review of our manuscript, Mr. Yabing Liu and Ms. Jianxia Peng for their expert technical assistance, and Ms. Qinghuan Li, for her excellent laboratory management. This work was supported by grants from the National Natural Science Foundation for Distinguished Young Scholars (C03020504, Y.Z.), the National Natural Science Foundation for Key Programs (No. 30630060, Y.Z.), the Knowledge Innovation Program of CAS (KSCX2-SW-333, Y.Z.), the National Basic Research Program (973 Program, 2003CB515501, Y.Z.), and the Scientific Research Foundation for the Returned Overseas Chinese Scholars of State Education Ministry (2005-546, Y.Z.).
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