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Heat stable ssDNA/RNA-binding activity of a wheat cold shock domain protein
Authors:Nakaminami Kentaro  Sasaki Kentaro  Kajita Shinya  Takeda Hisashi  Karlson Dale  Ohgi Kazuko  Imai Ryozo
Institution:Winter Stress Laboratory, Department of Low Temperature Sciences, National Agricultural Research Center for Hokkaido Region, NARO, Hitsujigaoka 1, Toyohira-ku, Sapporo 062-8555, Japan.
Abstract:The cold-induced wheat WCSP1 protein belongs to the cold shock domain (CSD) protein family. In prokaryotes and eukaryotes, the CSD functions as a nucleic acid-binding domain. Here, we demonstrated that purified recombinant WCSP1 is boiling soluble and binds ss/dsDNA and mRNA. Furthermore, boiled-WCSP1 retained its characteristic nucleic acid-binding activity. A WCSP1 deletion mutant, containing only a CSD, lost ssDNA/RNA-binding activity; while a mutant containing the CSD and the first glycine-rich region (GR) displayed the activity. These data indicated that the first GR of WCSP1 is necessary for the binding activity but is not for the heat stability of the protein.
Keywords:CD  circular dichroism  CSD  cold shock domain  GR  glycine-rich region  GST  glutathione-S-transferase  ZnF  zinc finger
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