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CELL BIOLOGY AND SIGNAL TRANSDUCTION

The Fertilization-Induced DNA Replication Factor MCM6 of Maize Shuttles between Cytoplasm and Nucleus, and Is Essential for Plant Growth and Development¹

Thomas Dresselhaus^*, Kanok-orn Srilunchang, Dunja Leljak-Levani, Daniela N. Schreiber² and Preeti Garg

Developmental Biology and Biotechnology, Biocenter Klein Flottbek, University of Hamburg, 22609 Hamburg, Germany (T.D., K.S., P.G.); and Department of Molecular Biology, Faculty of Science, University of Zagreb, 10000 Zagreb, Croatia (D.L.-L.)

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.

² Present address: Department of Biology, National University of Ireland, Maynooth, County Kildare, Ireland.

The author responsible for distribution of materials integral to the findings presented in this article in accordance with the policy described in the Instructions for Authors (www.plantphysiol.org) is: Thomas Dresselhaus (dresselh{at}botanik.uni-hamburg.de).

Article, publication date, and citation information can be found at www.plantphysiol.org/cgi/doi/10.1104/pp.105.074294.

^* Corresponding author; e-mail dresselh{at}botanik.uni-hamburg.de; fax 49–40–42816–229.

Received November 17, 2005; returned for revision December 8, 2005; accepted December 11, 2005.

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