Dresselhaus Thomas, Srilunchang Kanok-Orn, Leljak-Levanic Dunja, Schreiber Daniela N, Garg Preeti
Developmental Biology and Biotechnology, Biocenter Klein Flottbek, University of Hamburg, 22609 Hamburg, Germany.
Plant Physiol. 2006 Feb;140(2):512-27. doi: 10.1104/pp.105.074294. Epub 2006 Jan 11.
The eukaryotic genome is duplicated exactly once per cell division cycle. A strategy that limits every replication origin to a single initiation event is tightly regulated by a multiprotein complex, which involves at least 20 protein factors. A key player in this regulation is the evolutionary conserved hexameric MCM2-7 complex. From maize (Zea mays) zygotes, we have cloned MCM6 and characterized this essential gene in more detail. Shortly after fertilization, expression of ZmMCM6 is strongly induced. During progression of zygote and proembryo development, ZmMCM6 transcript amounts decrease and are low in vegetative tissues, where expression is restricted to tissues containing proliferating cells. The highest protein amounts are detectable about 6 to 20 d after fertilization in developing kernels. Subcellular localization studies revealed that MCM6 protein shuttles between cytoplasm and nucleoplasm in a cell cycle-dependent manner. ZmMCM6 is taken up by the nucleus during G1 phase and the highest protein levels were observed during late G1/S phase. ZmMCM6 is excluded from the nucleus during late S, G2, and mitosis. Transgenic maize was generated to overexpress and down-regulate ZmMCM6. Plants displaying minor antisense transcript amounts were reduced in size and did not develop cobs to maturity. Down-regulation of ZmMCM6 gene activity seems also to affect pollen development because antisense transgenes could not be propagated via pollen to wild-type plants. In summary, the transgenic data indicate that MCM6 is essential for both vegetative as well as reproductive growth and development in plants.
真核生物基因组在每个细胞分裂周期中精确复制一次。一种将每个复制起点限制为单个起始事件的策略受到一种多蛋白复合物的严格调控,该复合物至少涉及20种蛋白质因子。这种调控中的一个关键参与者是进化保守的六聚体MCM2 - 7复合物。我们从玉米(Zea mays)合子中克隆了MCM6,并对这个必需基因进行了更详细的表征。受精后不久,ZmMCM6的表达被强烈诱导。在合子和原胚发育过程中,ZmMCM6转录本数量减少,在营养组织中含量较低,其表达仅限于含有增殖细胞的组织。在发育中的籽粒中,受精后约6至20天可检测到最高的蛋白质含量。亚细胞定位研究表明,MCM6蛋白以细胞周期依赖性方式在细胞质和核质之间穿梭。ZmMCM6在G1期被细胞核摄取,在G1/S晚期观察到最高的蛋白质水平。在S期晚期、G2期和有丝分裂期间,ZmMCM6被排除在细胞核外。通过转基因玉米来过量表达和下调ZmMCM6。显示少量反义转录本的植株尺寸减小,并且未发育成熟的果穗。ZmMCM6基因活性的下调似乎也影响花粉发育,因为反义转基因不能通过花粉传播到野生型植株。总之,转基因数据表明MCM6对于植物的营养生长以及生殖生长和发育都是必需的。