9-1 copyright 2005 mcgraw-hill australia pty ltd ppts t/a biology: an australian focus 3e by knox,...
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![Page 1: 9-1 Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint Chapter 9: Genes, chromosomes](https://reader037.vdocuments.us/reader037/viewer/2022110322/56649d265503460f949fcbea/html5/thumbnails/1.jpg)
9-1Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Chapter 9: Genes, chromosomes and DNA
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9-2Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Tracking the genetic material
• 1869—chromatin isolated by Miescher, containing nucleic acid and protein
• Chromosomes consist of DNA and proteins• 1900—concept of ‘Mendelian inheritance’
controlled by ‘genes’• 1910—Morgan and others noted parallel
inheritance of ‘genes’ with chromosomes, suggesting that genes were ‘on’ the chromosomes
(cont.)
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9-3Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Tracking the genetic material (cont.)• The transforming principle in Streptococcus
pneumoniae, where virulence can be transferred by cellular extracts containing DNA (Avery, McLeod & McCarty 1944)
– mice injected with live non-virulent bacteria and heat-killed virulent bacterial material died
– neither preparation on its own killed the mice– non-virulent strain was ‘transformed’ by the virulent
material– the virulence acquired from the heat-killed strain was
passed on to progeny of the transformed bacteria
(cont.)
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9-4Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.5: Transforming principle in Streptococcus pneumoniae
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9-5Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Tracking the genetic material (cont.)• DNA, not protein, is the genetic information
(Hershey & Chase 1952)– bacteriophage DNA or protein was specifically
radioactively labelled– bacteriophage infected bacteria—new bacteriophage
produced by infected organisms– the presence of radiolabel inside infected bacteria was
only detected when the DNA was radiolabelled– no radiolabelled protein was found inside the bacteria
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9-6Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.6: Radioactive labelling of DNA with 32P or protein with 35S
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9-7Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Chromosomes
• DNA is organised into chromosomes• Each chromosome is a single DNA molecule• In eukaryotic cells, chromosomes are located in
the nucleus• Each species has a unique chromosome
complement—shape, size and number• Centromere essential for segregation during cell
division
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9-8Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.1: Stained human chromosomes
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9-9Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Chromosome structure
• Multiple levels of DNA folding– nucleosome: 146 base pairs (bp) are coiled in 1.75
turns around a core of histone proteins (H2A, H2B, H3, H4) 10 nm diameter
(cont.)
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9-10Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.3: Model of a nucleosome particle
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9-11Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Chromosome structure (cont.)
• This string of nucleosome ‘beads’ is then further coiled into chromatin fibres 30 nm diameter
• Metaphase chromosomes are further condensed to about 1/10 000 of their full length
• Loops of 20–100 kb are attached to a central protein scaffold
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9-12Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.4: A condensed chromosome in metaphase
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9-13Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
DNA structure
• DNA is a double-stranded molecule twisted into a helix
• Each strand, comprising a sugar-phosphate backbone and attached bases, is connected to a complementary strand by non-covalent hydrogen bonding between paired bases
• The bases are adenine (A), thymine (T), cytosine (C) and guanine (G)
(cont.)
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9-14Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
DNA structure (cont.)
• DNA consists of four different nucleotides• Each nucleotide has three parts: a phosphate
group, a pentose sugar and an organic base
(cont.)
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9-15Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.7: Molecular structure of DNA
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9-16Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
DNA structure (cont.)
• Bases are purines (A and G) and pyrimidines (C and T)
• Purines have a pair of fused rings; pyrimidines only have one
• A and T are connected by two hydrogen bonds; G and C are connected by three hydrogen bonds
• The number of bonds is the basis of specific pairing between the bases
(cont.)
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9-17Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
DNA structure (cont.)
• Nucleotides are linked together by phosphodiester bonds
• Nucleic acids have distinct ends – the 3’ end has a free hydroxyl group on the 3’ carbon of a
sugar – the 5’ end has a free phosphate group at the 5’ carbon of
the sugar
• The two strands of the helix are antiparallel: the 5’ end of one strand is directly apposed to the 3’ end of the other strand
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9-18Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
DNA replication
• DNA is replicated semi-conservatively—each separate strand provides the template for new strand synthesis by the base-pairing rules
• Semi-conservative replication allows synthesis of new strands with high fidelity
• New DNA molecules consist of one ‘old’ strand from the original molecule and one newly synthesised strand
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9-19Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.8a: Semiconservative replication
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9-20Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.8b: Sequence-based representation of replicating DNA
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9-21Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
DNA replication in prokaryotes
• Bacteria have a single circular chromosome• Replication begins at a single origin of replication• A nick is made in at least one strand and the
molecule unwinds• A replication fork is formed on each side of the
origin as small lengths of DNA separate for synthesis of new strands
• The two replication forks eventually meet at the terminus
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9-22Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.10: DNA synthesis in circular chromosomes
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9-23Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Enzymes in replication
• Requires gyrases to unwind the supercoiled helices and helicases to separate the strands
• New strand synthesis is performed by DNA polymerases
– DNA polymerase III attaches bases in the 5’ 3’ direction– DNA polymerase I checks the added base and corrects it
by 3’ to 5’ exonuclease activity—also removes RNA primers used to initiate replication
• DNA polymerases require priming to initiate strand extension
– a short RNA primer with a 3’ OH group is added to the template strand by a primase (cont.)
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9-24Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.13: Initiation of DNA synthesis
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9-25Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Enzymes in replication (cont.)
• Synthesis always proceeds 5’ 3’ on the strand being produced therefore
– one strand is synthesised continuously (leading strand)– the other (lagging strand) is synthesised discontinuously
as the replication fork moves along the template strand – primases attach a series of primers along the template
strand– DNA polymerase extends the primers away from the
replication fork– the resulting Okazaki fragments are then ligated by DNA
ligase
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9-26Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.11: Replication fork of Escherichia coli
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9-27Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Replication in eukaryotes
• Chromosomes have many origins of replication• Two replication forks are formed at each origin• Synthesis proceeds 5’ to 3’ at each unit of
replication (replicon) with leading and lagging strands
(cont.)
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9-28Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.14: DNA synthesis in a chromosome of a eukaryote
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9-29Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Replication in eukaryotes (cont.)
• Okazaki fragments are shorter than in prokaryotes• Leading and lagging strand synthesis in human
cells is performed by different DNA polymerases• Multiple replicons are necessary due to the large
size of eukaryote chromosomes• Replicons are initiated at different times
– chromosomes have early-, mid- or late-replicating regions
– gene-rich regions tend to be replicated first
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9-30Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Telomeres during replication
• DNA polymerases only replicate DNA 5’ to 3’ and need a primer
• When the primer is removed from the 5’ end of the new strand a gap is left from which DNA polymerase cannot extend
• At each round of cell division chromosomes would become shorter
(cont.)
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9-31Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Telomeres during replication (cont.)
• To overcome this problem– chromosomes have telomeres repeat DNA sequences up
to 10–15 kb– added to chromosome ends by telomerase– priming provided by RNA molecule within the telomerase
complex– chromosome length is maintained
(cont.)
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9-32Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Fig. 9.15: Completion of replication at ends (telomeres) of eukaryotic chromosomes
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9-33Copyright 2005 McGraw-Hill Australia Pty Ltd PPTs t/a Biology: An Australian focus 3e by Knox, Ladiges, Evans and Saint
Telomeres during replication (cont.)• Mammalian somatic cells have no telomerase
activity so become shorter with age• This limits the number of divisions each cell can
undergo• Essential sequences are eventually lost and the
cell dies• Restoration of telomerase activity allows cells to
proliferate indefinitely• Telomerase is important in ageing and cancer