Carter C, Graham R A, Thornburg R W
Department of Biochemistry and Biophysics, Iowa State University Ames 50011, USA.
Plant Mol Biol. 1998 Dec;38(6):929-43. doi: 10.1023/a:1006038117130.
We have identified 39 Arabidopsis thaliana ESTs encoding germin-like proteins (GLPs) and have completely sequenced 25 of these cDNAs. Our analysis demonstrates that the Arabidopsis genome contains a gene family with at least 12 GLP genes. Comparisons with other known germins and germin-like proteins indicate that these Arabidopsis GLP subfamilies are unique from wheat germin. All other known GLPs fall into one of these subfamilies. The translated GLPs show approximately 35% amino acid identity with other GLPs outside of their subfamily and significantly higher levels of identity within their respective subfamily. The 3' ends of many of the GLP cDNAs are heterogeneous and several sites of polyadenylation are used. Ten of the GLPs have N-terminal signal sequences and most appear to be exported from the cell. Structurally, the GLPs are predicted to have a high content of beta-pleated sheet. Seven conserved regions of beta-sheet were found in each of the GLP proteins along with alpha-helices located at both N- and C-termini. These same structural elements are also conserved in wheat germin. With one exception, all GLP family members contain at least one N-glycosylation site. All of these sites are conserved in an unstructured loop between beta-1 and beta-2. Genes for two of these GLPs were identified in genomic sequences previously deposited in the GenBank. The GLP3b gene is physically linked to the polyubiquitin 4 gene. The 3' end of the GLP3b mRNA is only 0.5 kb from the ubq4 start of transcription. Analysis of the GLP3b promoter shows the presence of a single putative auxin-response sequence located at -124 to -111 upstream from the 5' end of the GLP3b mRNA. The GLP9 gene was identified in an Arabidopsis contig from Chromosome 4.
我们已鉴定出39个编码类萌发素蛋白(GLP)的拟南芥EST,并对其中25个cDNA进行了全序列测定。我们的分析表明,拟南芥基因组包含一个至少有12个GLP基因的基因家族。与其他已知的萌发素和类萌发素蛋白比较显示,这些拟南芥GLP亚家族与小麦萌发素不同。所有其他已知的GLP都属于这些亚家族之一。翻译后的GLP与亚家族外的其他GLP显示约35%的氨基酸同一性,而在各自亚家族内的同一性水平明显更高。许多GLP cDNA的3'端是异质的,并且使用了多个聚腺苷酸化位点。其中10个GLP具有N端信号序列,大多数似乎从细胞中输出。在结构上,预测GLP具有高含量的β折叠片层。在每个GLP蛋白中发现了7个保守的β折叠区域,以及位于N端和C端的α螺旋。这些相同的结构元件在小麦萌发素中也保守。除了一个例外,所有GLP家族成员都至少含有一个N-糖基化位点。所有这些位点在β-1和β-2之间的无结构环中保守。在先前存入GenBank的基因组序列中鉴定出了其中两个GLP的基因。GLP3b基因与多聚泛素4基因物理连锁。GLP3b mRNA的3'端距离ubq4转录起始点仅0.5 kb。对GLP3b启动子的分析表明,在GLP3b mRNA 5'端上游-124至-111处存在一个单一的假定生长素反应序列。GLP9基因是在来自第4号染色体的拟南芥重叠群中鉴定出来的。