engleski

The fertilization-Induced DNA Replication Factor MCM6 of Maize Shuttles between Cytoplasm and Nucleus, and is Essential for Plant Growth and Development

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 multi-protein complex, which involves at least 20 protein factors. A key player in this regulation is the evolutionary conserved hexameric MCM2-7 complex. From Zea mays (L.) 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 pro-embryo development, ZmMCM6 transcript amounts decrease and are low in vegetative tissues, where expression is restricted to tissues containing proliferating cells. Highest protein amounts are detectable about 6-20 days 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 highest protein levels were observed during late G1/S phase. During G2 phase and mitosis, ZmMCM6 is excluded from the nucleus. Transgenic maize was generated to over-express and to 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 to affect also pollen development, as 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.

Fertilization; zygote; ZGA; cell cycle; maize

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