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Chapter 5 DNA and Chromosomes DNA as the genetic material Heat-killed bacteria can transform living cells S Smooth R Rough Fred Griffith, 1920 DNA is the genetic material Oswald Avery Colin MacLeod Maclyn McCarty 1935 1. The first evidence that DNA could serve as the genetic material 2. The genetic material was likely to be made of protein Genes are made of DNA Martha Chase Alfred Hershey 1952 The empty viral coats T2 virus Radioactively label (w/o S) (w/o P) The Nobel Prize in Physiology or Medicine 1969 The replication mechanism and the genetic structure of viruses What is gene? Genes – the information-containing elements that determine the characteristics of a species as a whole and of the individuals within it. A gene is usually defined as a segment of DNA that contains the instructions for making a particular protein (or, in some cases, a set of closely related proteins). What is genome? Genome – the totality of DNA genetic information in each cell is called its genome. = 3.2x109 nucleotides. What is chromosome? Chromosomes – in eucaryotic cells, very long double-strand DNA molecules are packaged into structure called chromosomes. Homologous chromosomes (homologs) Homologus chromosomes – the maternal and paternal chromosomes of a pair. Chromosomes become visible as cells prepare to divide DAPI DNA staining Nucleotide DNA is made of four nucleotide building blocks Sugar-phosphate backbone Polynucleotide chain Antiparallel The two strands of the DNA double helix are hold together by hydrogen bonds between complementary base pairs Base pair Antiparallel 5’-PO4 3’-OH 5’-PO4 3’-OH A space-filling model shows the conformation of DNA double helix 10 bases/turn P O H N Genetic code Genetic code – the exact correspondence between the 4-letter nucleotide alphabet of DNA and the 20-letter amino acid alphabet of proteins Transcription and translation Linear messages come in many forms Gene contain information to make proteins Each human chromosome can be “painted” a different color to allow its unambiguous identification under the light microscope DNA hybridization Karyotype Karyotypes - The homologus chromosomes are numbered and arranged in pairs Unique banding patterns allow the identification of the human chromosomes Giemsa stain A-T rich centromere rRNAs centromere Abnormal chromosomes are associated with some inherited genetic defects Ataxia / Chromosome 12 Chromosome 4 Genes are arranged along the chromosomes Junk DNA Gene Closely related species can have very different chromosome numbers The two species shown have roughly the same number of genes The cell cycle The replication and segregation of chromosomes occurs through an ordered cell cycle in proliferating cells microtubules G0/G1 S G2 M Three DNA sequence elements are needed to produce a eucaryotic chromosome that can be replicated then segregation at mitosis x2 x1 Replication origin DNA Replication Centromere The role of telomere in life span A typical mitotic chromosome is highly compact Mitotic chromosome SEM Interphase chromosomes occupy different territories within the nucleus The nucleolus is the most prominent structure in the interphase nucleus (Chromosome13, 14, 15, 21, 22) TEM Nucleolus Nucleolus – The parts of the different chromosomes carrying genes for ribosomal RNA (rRNA) cluster together. Heterochromatin & Euchromatin Heterochromatin – The most highly condensed form of interpahse chromatin is called heterochromatin. DNA in interphase chromosome is less compact than that in mitotic chromosomes Interphase nuclear DNA Mitotic chromosome TEM Nucleosomes can be seen in the electron microscope Chromatin 30-nm thick Unpacked, decondensed “beads-on-a-string” Nucleosome Nucleosome core particle DNA TEM Chromatin Chromatin – The complex of both classes of protein with nuclear DNA. Most of the chromatin is in the form of fibers, each with a diameter of about 30 nm. Nucleosome Nucleosome – The first and most fundamental level of chromatin packing. Histones Nucleosomes contain DNA wrapped around a protein core of eight histone molecules High proportion of positively charged amino acids (lysine and arginine) 80bp Negatively-charged sugar-phosphate backbone of DNA The structure of the nucleosome core particle, as determined by X-ray diffraction analysis, reveals how DNA is tightly wrapped around a disc-shaped histone core Highly conserved 1.7 turns in a left-handed coil N’ N’ A linker histone helps to pull nucleosomes together into the 30-nm fiber DNA packing occurs on several levels in chromosomes Interphase chromatin Mitotic chromasome + Histone H1 The mitotic chromosome is formed from tightly packed chromatin SEM Chromosome & Chromatids Chromatin-remodeling complexes reposition the DNA wrapped around nucleosomes DNA-binding proteins The pattern of modification of histone tails can dictate how a stretch of chromatin is treated by the cell To bind specific proteins N’ Methyl group (+) Acetyl group (-) Phosphate (-) Interphase chromatin Heterochromatin: 10% Euchromatin: 90% Expression of a gene can be altered by moving it to another location in the genome (expression) Position effect (non-expression) An X chromosome can be inactivated by heterochromatin formation Double dose of X-chromosome prodicts would be lethal The structure of chromatin varies along a single interphase chromosome How histone modifications may be inherited by daughter chromosomes Restore the parental modification Epigenetic inheritance Epigenetics – The study of heritable changes in gene expression or cellular phenotype caused by mechanisms other than changes in the underlying DNA sequence.