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Golgi localization of Syne-1

机译：Syne-1的高尔基体定位

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We have previously identified a Golgi-localized spectrin isoform by using an anti-body to the beta-subunit of erythrocyte spectrin. In this study, we show that a screen of a lambdagt11 expression library resulted in the isolation of an similar to5-kb partial cDNA from a Madin-Darby bovine kidney (MDBK) cell line, which encoded a polypeptide of 1697 amino acids with low, but detectable, sequence homology to spectrin (37%). A blast search revealed that this clone overlaps with the 5' end of a recently identified spectrin family member Syne-1B/Nesprin-1beta, an alternately transcribed gene with muscle-specific forms that bind acetylcholine receptor and associate with the nuclear envelope. By comparing the sequence of the MDBK clone with sequence data from the human genome database, we have determined that this cDNA represents a central portion of a very large gene (similar to500 kb), encoding an similar to25-kb transcript that we refer to as Syne-1. Syne-1 encodes a large polypeptide (8406 amino acids) with multiple spectrin repeats and a region at its amino terminus with high homology to the actin binding domains of conventional spectrins. Golgi localization for this spectrin-like protein was demonstrated by expression of epitope-tagged fragments in MDBK and COS cells, identifying two distinct Golgi binding sites, and by immunofluorescence microscopy by using several different antibody preparations. One of the Golgi binding domains on Syne-1 acts as a dominant negative inhibitor that alters the structure of the Golgi complex, which collapses into a condensed structure near the centrosome in transfected epithelial. cells. We conclude that the Syne-1, gene is, expressed in a variety of forms that are multifunctional and are capable of functioning at both the Golgi and the nuclear envelope, perhaps linking the two organelles during muscle differentiation. [References: 34]
机译：我们以前已经通过使用针对红细胞血影蛋白β亚基的抗体鉴定了高尔基体定位的血影蛋白同工型。在这项研究中，我们发现lambdagt11表达文库的筛选导致从Madin-Darby牛肾（MDBK）细胞系中分离出类似的5kb部分cDNA，该细胞系编码具有1697个氨基酸的低，但可检测到与血影蛋白的序列同源性（37％）。爆炸搜索显示该克隆与最近鉴定出的血影蛋白家族成员Syne-1B / Nesprin-1beta的5'末端重叠，后者是一种交替转录的基因，具有结合乙酰胆碱受体并与核膜结合的肌肉特异性形式。通过将MDBK克隆的序列与人类基因组数据库的序列数据进行比较，我们已经确定该cDNA代表了一个非常大的基因（类似于500 kb）的中心部分，编码了一个类似于25 kb的转录本，我们称之为Syne-1。 Syne-1编码具有多个血影蛋白重复序列​​的大多肽（8406个氨基酸），并且在其氨基末端具有与常规血影蛋白的肌动蛋白结合域高度同源的区域。通过在MDBK和COS细胞中表位标记的片段表达，鉴定两个不同的高尔基体结合位点以及通过使用几种不同的抗体制剂进行的免疫荧光显微镜检查，证明了这种血影蛋白样蛋白的高尔基体定位。 Syne-1上的一个高尔基体结合结构域之一是一个主要的负性抑制剂，它改变了高尔基体的结构，该复合体在转染的上皮细胞的中心体附近塌陷成浓缩结构。细胞。我们得出的结论是，Syne-1基因以多种形式表达，这些形式是多功能的，并且能够在高尔基体和核膜上起作用，可能在肌肉分化过程中将两个细胞器联系在一起。 [参考：34]

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