Why study DNA replication?

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Presentation transcript:

Why study DNA replication? Attractive model for multicellular eukaryotes Small, sequenced genomes Genomic tools available Rapidly dividing cell cultures Tractable developmental programs Arabidopsis Rice Intercalary meristem Leaf blade Leaf sheath

Why study DNA replication? Potential practical applications Genetic engineering of artificial chromosomes Manipulating the endocycle New strategies for geminivirus resistance Cotton leaf curl disease Tomato leaf curl disease

Experimental Design One hour BrdU Pulse Fix Cells and Isolate Nuclei FACS “synchronization” IP BrdU-labeled DNA Label IP and Input DNA with Cy3/5 Hybridize to Chr4 Tiling Array 20000 probes 1 kb PCR products 95% coverage Normalize Smooth

Arabidopsis Chromosome 4 genes transposable elements nucleolar organizing region centromere

Replication Timing Profile log (early/late) L chromosome coordinate

Origin assembly Pre-replication complex licensing Initiation complexes assemble in late M and early G1 ORC - origin recognition complex MCM - replicative DNA helicase CDT1/CDC6 - MCM loader licensing

MCM5/7 are released during mitosis Merge DAPI DIC MCM5 MCM7 N. tabacum cultured cells (Nt-1)

How is licensing regulated in plants? X CDT1 Proteosome CDKA P A different CDT regulatory protein? Geminin ORC1 regulation? MCM subunits X Other mechanisms? cytoplasm nucleus

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Genetics Kit Activities Heredity & Genetics Scavenger Hunt Dr. Jeff Batten - Overview science activities Genetics Kit Activities Heredity & Genetics Scavenger Hunt Personal Survey of Inherited Human Traits Modeling Mendel Punnett Squares Genetic Offspring Models Extracting DNA from Wheat Germ DNA, Genes, and Proteins (Sickle Cell Disease) A Day in the Life of DNA Genetic Disorders Genetic vs. Environmental Influences on Traits

Activity 2: Survey of Inherited Traits Activity 5: Genetic Offspring Models Activity 6: Extracting Wheat DNA Activity 8: Day in the Life of DNA

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