Golgi localization of Syne-1

Lisa Lucio Gough, Jun Fan, Stephen Chu, Shawn Winnick, Kenneth A Beck

Research output: Contribution to journalArticle

61 Scopus citations

Abstract

We have previously identified a Golgi-localized spectrin isoform by using an antibody to the β-subunit of erythrocyte spectrin. In this study, we show that a screen of a λgt11 expression library resulted in the isolation of an ∼5-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-1β, 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 (∼500 kb), encoding an ∼25-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.

Original languageEnglish (US)
Pages (from-to)2410-2424
Number of pages15
JournalMolecular Biology of the Cell
Volume14
Issue number6
DOIs
StatePublished - Jun 1 2003

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics
  • Cell Biology

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    Gough, L. L., Fan, J., Chu, S., Winnick, S., & Beck, K. A. (2003). Golgi localization of Syne-1. Molecular Biology of the Cell, 14(6), 2410-2424. https://doi.org/10.1091/mbc.E02-07-0446