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Organization of Chromosomes--Study Guide and Outline Broad course objective: a.) explain the molecular structure of chromosomes as it relates to DNA packaging, chromosome function and gene expression Necessary for future material on: Chromosome Variation, Regulation of Gene Expression DNA Packaging—Why and How • If the DNA in a typical human cell were stretched out, what length would it be? What is the diameter of the nucleus in which human DNA must be packaged? • What degree of DNA packaging corresponds with “diffuse DNA” associated with G1? What kind of DNA packaging is associated with Mphase (“condensed DNA”)? • What types of DNA sequences make up the genome? What functions do they serve? • What are the differences between euchromatin and heterochromatin? • What types of proteins are involved in chromosome packaging? – – How do nucleosomes and histone proteins function in DNA packaging? What is chromosome scaffolding? How much DNA do different organisms have? Organism T4 Bacteriophage HIV E. colibacteria Yeast haploid genome in bp 168,900 9,750 4,639,221 13,105,020 Lily 36,000,000,000 Amoeba 290,000,000,000 Frog 3,100,000,000 Human 3,400,000,000 DNA content does not directly coincide with complexity of the organism. Any theories on why? Size measurements in the molecular world • 1 mm (millimeter) = 1/1,000 meter • 1 mm (“micron”) = 1/1,000,000 of a meter (1 x 10-6) • 1 nm (nanometer) = 1 x 10-9 meter •1 bp (base pair) = 1 nt (nucleotide pair) •1,000 bp = 1 kb (kilobase) •1 million bp = 1 Mb (megabase) •5 billion bp DNA ~ 1 meter •5 thousand bp DNA ~ 1.2 mm Representative genome sizes • Phage virus: 168 kb 65 nm phage head (~1,000 x length) • E. coli bacteria: 1,100 mm DNA ~0.2 micron space nucleoid region (5,500 x) • Human cell: 7.5 feet of DNA ~3 micron nucleus (2.3 million times longer than the nucleus) DNA packaging: How does all that DNA fit into one nucleus? (Equivalent to fitting 690 miles of movie film into a 30-foot room) An organism’s task in managing its DNA: 1.) Efficient packaging and storage, to fit into very small spaces (2.3 million times smaller) 2.) Requires “de-packaging” of DNA to access correct genes at the correct time (gene expression). 3.) Accurate DNA replication during the Sphase of the cell-cycle. Chromosomal puffs in condensed Drosophila chromosome show states of de-condensing in expressed regions Prokaryotic genome characteristics 1. Circular chromosome (only one), not linear 2. Efficient—more gene DNA, less or no Junk DNA 3. One origin sequence per chromosome How does the bacterial chromosome remain in its “tight” nucleoid without a nuclear membrane? Bacterial chromosome is normally supercoiled (~ 40 kb) Bacterial DNA released from supercoiling Bacterial DNA Supercoiling Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. Levels of DNA Packaging in Eukaryotes Types of DNA sequences making up the eukaryotic genome DNA type Unique-sequence Function Number/genome Protein coding and non-coding 1 Repetitive-sequence Opportunistic? Centromere Cytoskeleton attachment Telomere C’some stability few-107 1 region/c’some Ends of c’some DNA Centromere sequences •Repeating sequences •Non protein-coding •Sequences bind to centromere proteins, provide anchor sites for spindle fibers Reminder of function of kinetochores and kinetochore microtubules Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. Chromosome fragments lacking centromeres are lost in mitosis (Figure 11.10) Telomere sequences function to preserve the length of the “ends” Dolly: First successful cloned adult animal Born on July 5, 1996, Dolly died on February 14, 2003. Dolly suffered from lung disease, heart disease and other symptoms of premature aging. Telomeres sequences may loop back and preserve DNA-ends during replication Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. Major proteins necessary for chromosome structure Protein type Function Histone packaging at 11nm width, nucleosome formation Linker proteins packaging at 11nm width, nucleosome formation Scaffold “Skeleton” of the condensed mitotic c’some Kinetochore Cytoskeleton attachment to centromere Telomerase enzyme for preserving lengths of telomeres in stem cells (covered in DNA Replication chapter) Telomere caps degradation protects ends of linear chromosomes from Levels of DNA Packaging in Eukaryotes Digestion of nucleosomes reveals nucleosome structure Nucleosome structure Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. •Positively charged histone “tails” bind to DNA. •Acetylation of histone proteins allows access to DNA COCH3-- -COCH3 -COCH3 Transcription Factor Levels of DNA Packaging in Eukaryotes Arrangement of 30-nm chromatin fiber into looped domains Peter J. Russell, iGenetics: Copyright © Pearson Education, Inc., publishing as Benjamin Cummings. Eukaryotic DNA “released” from its tight packaging as a metaphase chromosome (only scaffold remains) Go over lecture outline at end of lecture