Bark I C
Department of Developmental Biology, Uppsala University, Biomedical Center, Sweden.
J Mol Biol. 1993 Sep 5;233(1):67-76. doi: 10.1006/jmbi.1993.1485.
SNAP-25 (synaptosomal associated protein of 25 kDa) is a neuronal-specific but non-uniformly expressed protein. Expression is correlated to the time of synaptogenesis and SNAP-25 protein is found predominantly in presynaptic fields. Here we report the cloning and characterization of the SNAP-25 chicken gene. The gene spans more than 65 kb of genomic DNA and is interrupted into nine different exons. Identification of the 5'-end of the mRNA showed that transcriptional initiation is heterogeneous, although a major transcriptional start site is present 30 bp downstream of a putative TATA-box. Analysis of the exon-intron organization revealed that exon 5 exists in two homologous but distinct versions separated by 140 base-pairs. Both exons 5 are expressed as mRNA, indicating that alternative splicing regulates the expression of two isoforms, a and b, of the SNAP-25 protein. The alternative splicing results in a difference of nine amino acid residues between the proteins in a domain demonstrated to be subject to palmitoylation. This implies that the two isoforms differ in their capacity or efficiency to be modified by fatty acylation, suggesting divergent abilities to interact with neuronal membranes.
SNAP-25(25 kDa突触体相关蛋白)是一种神经元特异性但表达不均一的蛋白质。其表达与突触发生时间相关,且SNAP-25蛋白主要存在于突触前区域。在此,我们报告SNAP-25鸡基因的克隆与特征分析。该基因跨越超过65 kb的基因组DNA,被打断成九个不同的外显子。对mRNA 5'端的鉴定表明,转录起始是异质性的,尽管一个主要转录起始位点位于假定TATA框下游30 bp处。对外显子-内含子组织的分析显示,外显子5存在两个同源但不同的版本,相隔140个碱基对。两个外显子5均表达为mRNA,这表明可变剪接调节了SNAP-25蛋白两种异构体a和b的表达。可变剪接导致两种蛋白质在一个被证明可进行棕榈酰化的结构域中存在九个氨基酸残基的差异。这意味着这两种异构体在被脂肪酰化修饰的能力或效率上存在差异,表明它们与神经元膜相互作用的能力不同。