Marian Calin O, Bordoli Stefano J, Goltz Marion, Santarella Rachel A, Jackson Leisa P, Danilevskaya Olga, Beckstette Michael, Meeley Robert, Bass Hank W
Department of Biological Science, Florida State University, Tallahassee, Florida 32306-4370, USA.
Plant Physiol. 2003 Nov;133(3):1336-50. doi: 10.1104/pp.103.026856. Epub 2003 Oct 23.
We screened maize (Zea mays) cDNAs for sequences similar to the single myb-like DNA-binding domain of known telomeric complex proteins. We identified, cloned, and sequenced five full-length cDNAs representing a novel gene family, and we describe the analysis of one of them, the gene Single myb histone 1 (Smh1). The Smh1 gene encodes a small, basic protein with a unique triple motif structure of (a) an N-terminal SANT/myb-like domain of the homeodomain-like superfamily of 3-helical-bundle-fold proteins, (b) a central region with homology to the conserved H1 globular domain found in the linker histones H1/H5, and (c) a coiled-coil domain near the C terminus. The Smh-type genes are plant specific and include a gene family in Arabidopsis and the PcMYB1 gene of parsley (Petroselinum crispum) but are distinct from those (AtTRP1, AtTBP1, and OsRTBP1) recently shown to encode in vitro telomere-repeat DNA-binding activity. The Smh1 gene is expressed in leaf tissue and maps to chromosome 8 (bin 8.05), with a duplicate locus on chromosome 3 (bin 3.09). A recombinant full-length SMH1, rSMH1, was found by band-shift assays to bind double-stranded oligonucleotide probes with at least two internal tandem copies of the maize telomere repeat, TTTAGGG. Point mutations in the telomere repeat residues reduced or abolished the binding, whereas rSMH1 bound nonspecifically to single-stranded DNA probes. The two DNA-binding motifs in SMH proteins may provide a link between sequence recognition and chromatin dynamics and may function at telomeres or other sites in the nucleus.
我们筛选了玉米(Zea mays)cDNA,寻找与已知端粒复合体蛋白的单个类myb DNA结合结构域相似的序列。我们鉴定、克隆并测序了代表一个新基因家族的五个全长cDNA,并描述了其中一个基因——单myb组蛋白1(Smh1)基因的分析。Smh1基因编码一种小的碱性蛋白,具有独特的三重基序结构:(a)一个3-螺旋束折叠蛋白的同源异型域样超家族的N端SANT/类myb结构域;(b)一个与连接组蛋白H1/H5中保守的H1球状结构域具有同源性的中央区域;(c)靠近C端的一个卷曲螺旋结构域。Smh型基因是植物特有的,包括拟南芥中的一个基因家族以及欧芹(Petroselinum crispum)的PcMYB1基因,但与最近显示在体外编码端粒重复DNA结合活性的那些基因(AtTRP1、AtTBP1和OsRTBP1)不同。Smh1基因在叶片组织中表达,定位于第8号染色体(8.05区间),在第3号染色体上有一个重复位点(3.09区间)。通过凝胶迁移实验发现,重组全长SMH1(rSMH1)能与含有至少两个内部串联拷贝的玉米端粒重复序列TTTAGGG的双链寡核苷酸探针结合。端粒重复序列残基中的点突变会降低或消除这种结合,而rSMH1能非特异性地结合单链DNA探针。SMH蛋白中的两个DNA结合基序可能在序列识别和染色质动力学之间提供联系,并可能在端粒或细胞核中的其他位点发挥作用。