US-Czech Collaborative Research on Nuclear Myosin I

该奖项将支持美国和捷克合作研究最近发现的新型核肌球蛋白。 资金（由国际方案司提供）将严格用于促进和促进研究的国际部分。 来自捷克共和国布拉格的de Lanerolle博士和他的合作者Pavel Hozak博士已经证明了细胞核中存在一种类似肌球蛋白I的蛋白质。 该蛋白质的表观分子量为120，000，与K+-EDTA ATP酶活性相关，与钙调蛋白结合，用ATP进行光亲和标记，在不存在但不存在ATP的情况下与肌动蛋白结合，并被从肾上腺纯化的肌球蛋白I的抗体识别。 120 kDa蛋白的微测序已将其明确鉴定为基于肌动蛋白的分子马达的肌球蛋白超家族的肌球蛋白I-β亚家族的成员。 微测序数据用于克隆该蛋白的cDNA。 这两种方法都证明了120 kDa蛋白特有的16个氨基酸N-末端延伸的存在。 其他数据表明，独特的16个氨基酸的延伸是负责核肌球蛋白I的核定位和核肌球蛋白I与RNA聚合酶II，并参与转录。 de Lanerolle博士和Hozak博士现在建议继续这项国际合作，进一步探索这些发现。 共定位和功能测定将在布拉格的Hozak博士实验室进行，使用芝加哥的de Lanerolle博士实验室开发的质粒、抗体和其他试剂。 为了促进这种合作，Hozak博士的学生将在De Lanerolle博士的实验室接受分子生物学方法的培训，De Lanerolle博士的学生将在Hozak博士的实验室接受先进的显微镜和显微注射方法以及原位转录测定的培训。

This award will support a US-Czech collaboration on studies of a recently discovered novel nuclear myosin. The funds (provided from the Division of International Programs) will be used strictly to promote and facilitate the international component of the research. Dr. de Lanerolle and his collaborator, Dr. Pavel Hozak in Prague, Czech Republic have already demonstrated the presence of a myosin I like protein in the nucleus. This protein has an apparent molecular weight of 120,000, it is associated with K+-EDTA ATPase activity, it binds calmodulin, it is photoaffinity labelled with ATP, it binds actin in the absence, but not the presence, of ATP and it is recognized by an antibody to myosin I purified from adrenal glands. Microsequencing of the 120 kDa protein has positively identified it as a member of the myosin I-beta subfamily of the myosin superfamily of actin based molecular motors. The microsequencing data was used to clone the cDNA for this protein. Both methods have demonstrated the presence of a 16 amino acid N-terminal extension that is unique to the 120 kDa protein. Other data suggest that the unique 16 amino acid extension is responsible for the nuclear localization of nuclear myosin I and that nuclear myosin I associates with RNA polymerase II and is involved in transcription. Drs. de Lanerolle and Hozak now propose to continue this international collaboration to explore these findings further. Co-localization and functional assays will be performed in Dr. Hozak's laboratory in Prague, using plasmids, antibodies and other reagents developed in Dr. de Lanerolle's laboratory in Chicago. To facilitate this collaboration, Dr. Hozak's students will be trained in molecular biology methods in Dr. De Lanerolle's laboratory and Dr. de Lanerolle's students will be trained in advanced microscopy and microinjection methods and in situ transcription assays in Dr. Hozak's laboratory.