Titus M A, Novak K D, Hanes G P, Urioste A S
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Biophys J. 1995 Apr;68(4 Suppl):152S-155S; discussion 156S-157S.
Several new members of the Dictyostelium myosin family have been identified by physical mapping techniques in combination with PCR. Here we describe the initial molecular genetic characterization of one of these, myoF. A 1-kb segment of the myoF gene was obtained by the PCR and used as a specific probe for Northern analysis and as a vehicle for gene-targeting studies. The myoF gene is expressed as a 3.7-kb message, a size consistent with it encoding a myosin I class unconventional myosin, bringing the total of myosin is present in Dictyostelium to six. Analysis of strains in which the myoF gene has been disrupted reveals that loss of the myoF protein does not result in obvious defects either in cellular translocation, or in other readily assayed actin-based processes. The results of our investigation indicate that the myosin I family is quite large in Dictyostelium, and that several members, including myoF, may either be functionally redundant or play roles in as yet undescribed actin-based processes.
通过物理图谱技术结合聚合酶链反应（PCR），已鉴定出盘基网柄菌肌球蛋白家族的几个新成员。在此，我们描述其中一个成员——肌球蛋白F（myoF）的初步分子遗传学特征。通过PCR获得了myoF基因的一个1千碱基对（kb）片段，并将其用作Northern分析的特异性探针以及基因靶向研究的载体。myoF基因表达为一条3.7 kb的信使核糖核酸（mRNA），其大小与编码一种I类非传统肌球蛋白一致，这使得盘基网柄菌中存在的肌球蛋白总数达到六个。对myoF基因已被破坏的菌株进行分析发现，myoF蛋白的缺失在细胞迁移或其他易于检测的基于肌动蛋白的过程中均未导致明显缺陷。我们的研究结果表明，盘基网柄菌中的肌球蛋白I家族相当庞大，并且包括myoF在内的几个成员可能要么在功能上是冗余的，要么在尚未描述的基于肌动蛋白的过程中发挥作用。
