IDENTIFICATION AND OVERLAPPING EXPRESSION OF MULTIPLE UNCONVENTIONAL MYOSIN GENES IN VERTEBRATE CELL-TYPES
IDENTIFICATION AND OVERLAPPING EXPRESSION OF MULTIPLE UNCONVENTIONAL MYOSIN GENES IN VERTEBRATE CELL-TYPES
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DOI：
10.1073/pnas.91.14.6549
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发表时间：
1994-07-05
影响因子：
11.1
通讯作者：
MOOSEKER, MS
中科院分区：
文献类型：
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作者：
BEMENT, WM;HASSON, T;MOOSEKER, MS
分析了人上皮细胞系Caco-2（BBe）、猪上皮细胞系LLC-PK 1（CL-4）、人外周血白细胞和人肝脏中的肌球蛋白多样性。PCR扩增产生8-11个推定的肌球蛋白（取决于cDNA来源），代表六个不同的肌球蛋白类。通过杂交筛选获得的克隆的分析表明，原始PCR产物对应于真正的肌球蛋白，基于在其他肌球蛋白中高度保守的序列的存在。RNA酶保护分析证实了Caco-2（BBe）细胞中11种肌球蛋白的mRNA表达。免疫印迹分析表明，Caco-2（BBe）细胞至少表达6种肌球蛋白免疫原。结果显示，至少存在11个非常规的人类肌球蛋白基因，其中大多数在不同的细胞类型中以重叠的方式表达。肌球蛋白的丰度表明，肌球蛋白I与肌球蛋白II范例是不足以解释肌动蛋白为基础的细胞运动。
Myosin diversity in the human epithelial cell line Caco-2(BBe), the porcine epithelial cell line LLC-PK1 (CL-4), human peripheral blood leukocytes, and human liver was analyzed. PCR amplification yielded 8-11 putative myosins (depending on the cDNA source) representing six distinct myosin classes. Analysis of clones obtained by hybridization screening demonstrated that the original PCR products correspond to bona fide myosins, based on the presence of sequences highly conserved in other myosins. RNase protection analysis confirmed mRNA expression of 11 myosins in Caco-2(BBe) cells. Immunoblot analysis showed that at least 6 myosin immuno gens are expressed in Caco-2(BBe) cells. The results reveal the existence of at least 11 unconventional human myosin genes, most of which are expressed in an overlapping fashion in different cell types. The abundance of myosins suggests that the myosin I vs. myosin Il paradigm is inadequate to explain actin-based cellular motility.