武汉大学 分子生物学全部课程 3

1、Welcome Each of You to My Molecular Biology ClassMolecular Biology of the Gene, 5/E - Watson et al. (2004)Part I: Chemistry and Genetics Part II: Maintenance of the Genome Part III: Expression of the GenomePart IV: RegulationPart V: Methods3/08/05Part II: Maintenance of the GenomeDedicated to the st

2、ructure of DNA and the processes that propagate, maintain and alter it from one cell generation to the nextMaintenance of the GenomeCh 6: The structures of DNA and RNA Ch 7: Chromosomes, chromatins and the nucleosomeCh 8: The replication of DNACh 9: The mutability and repair of DNACh 10: Homologous

3、recombination at the molecular levelCh 11: Site-specific recombination and transposition of DNA3/08/05CHAPTER 6The Structures of DNA and RNA How do the structures of DNA and RNA account for their functions?OUTLINE1.DNA Structure2.DNA Topology3.RNA StructureDNA STRUCTUREDNA STRUCTURE (1) DNA is compo

4、sed of polynucleotide chains Structure: twisting around each other in the form of a double helix.Schematic modelSpace-filling modelNucleoside & Nucleotide, the fundamental building block of DNAglycosidic bondphosphoester bondDNA polarity: is defined by the asymmetry of the nucleotides and the wa

5、y they are joined.Phosphodiester linkages: repeating, sugar-phosphate backbone of the polynucleotide chainBases in DNApurinespyrimidinesadenineguaninecytosinethymineN9N1DNA STRUCTURE (2)Each bases has its preferred tautomeric form (Related to Ch 9)The two strands of the double helix are held togethe

6、r by base pairing in an antiparallel orientation, Which is a stereochemical consequence of the way that adenine and thymine,and guanine and cytosine, pair with each other. (Related to replication and transcription)DNA STRUCTURE (3)The Two Chains of the Double Helix Have Complementary SequencesExampl

7、e: If sequence 5-ATGTC-3 on one chain, the opposite chain must have the complementary sequence 3-TACAG-5DNA STRUCTURE (4)Watson-Crick Base Pairing(Related to replication and transcription)The strictness of the rules for “Waston-Crick” pairing derives from the complementarity both of shape and of hyd

8、rogen bonding properties between adenine and thymine and between guanine and cytosine.A:C incompatibilityHydrogen Bonding Is Important for the Specificity of Base PairingDNA STRUCTURE (5) The hydrogen bonds between complementary bases contribute to the thermodynamic stability of the helix (why?) and

9、 the specificity of base pairing Stacking interactions between bases significantly contribute to the stability of DNA double helixThe double helix has Minor and Major grooves (What & Why)DNA STRUCTURE (5)(See the Structural Tutorial of this chapter for details)It is a simple consequence of the g

10、eometry of the base pair.The Major groove is rich in chemical information (What are the biological relevance?)DNA STRUCTURE (6)The edges of each base pair are exposed in the major and minor grooves, creating a pattern of hydrogen bond donors and acceptors and of van der Waals surfaces that identifie

11、s the base pair.A: H-bond acceptorsD: H-bond donorsH: non-polar hydrogensM: methyl groupsThe double helix exists in multiple conformations.DNA STRUCTURE (7) The B form (10 bp/turn), which is observed at high humidity, most closely corresponds to the average structure of DNA under physiological condi

12、tionsA form (11 bp/turn), which observed under the condition of low humidity, presents in certain DNA/protein complexes. RNA double helix adopts a similar conformation. DNA strands can separate (denature) and reassociate (anneal)DNA STRUCTURE (8)Key terms to understand1.Denaturation2.Hybridization3.

13、Annealing/renature4.Absorbance 5.Hyperchromicity6.Tm (melting point)DNA TOPOLOGYDNA TOPOLOGY (1)Structure (1): Linking number is an invariant topological property of covalently closed, circular DNA (cccDNA)Linking number is the number of times one strand have to be passed through the other strand in

14、 order for the two strands to be entirely separated from each other.Species of cccDNA1. Plasmid and circular bacterial chromosomes 2. Linear DNA molecules of eukaryotic chromosomes due to their extreme length, entrainment in chromatin and interaction with other cellular components (Ch 7)Structure (2

15、): Linking number is composed of Twist and WritheThe linking number is the sum of the twist and the writhe.Twist is the number of times one strand completely wraps around the other strand.Writhe is the number of times that the long axis of the double helical DNA crosses over itself in 3-D space.DNA

16、TOPOLOGY (2)Local disruption of base pairsFunction (1): DNA in cells is negatively supercoiled; nucleosomes introduces negative supercoiling in eukaryotesNegative supercoils serve as a store of free energy that aids in processes that require strand separation, such as DNA replication and transcripti

17、on. Strand separation can be accomplished more easily in negatively supercoiled DNA than in relaxed DNADNA TOPOLOGY (3)Function (2): Topoisomerases (P115-119)1. The biological importance of topoisomerase?2. The functional difference of the two types of topoisomerases? 3. The working mechanism of top

18、oisomerase (See the animation for detail)DNA TOPOLOGY (4)RNA STRUCTURERNA STRUCTURE (1)RNA contains ribose and uracil and is usually single-strandedBiological roles of RNA1. RNA is the genetic material of some viruses2. RNA functions as the intermediate (mRNA) between the gene and the protein-synthe

19、sizing machinery.3. RNA functions as an adaptor (tRNA) between the codons in the mRNA and amino acids.4. RNA serves as a regulatory molecule, which through sequence complementarity binds to, and interferes with the translation of certain mRNAs.5. Some RNAs are enzymes that catalyze essential reactio

20、ns in the cell (RNase P ribozyme, large rRNA, self-splicing introns, etc).Structure (1): RNA chains fold back on themselves to form local regions of double helix similar to A-form DNARNA STRUCTURE (2)hairpinbulgeloopRNA helix are the base-paired segments between short stretches of complementary sequ

21、ences, which adopt one of the various stem-loop structuresSome tetraloop sequence can enhance the stability of the RNA helical structures For example, UUCG loop is unexpectedly stable due to the special base-stacking in the loopPseudoknots are complex structure resulted from base pairing of disconti

22、guous RNA segmentsFigure 6-32 Pseudoknot.Non-Watson-Crick G:U base pairs represent additional regular base pairing in RNA, which enriched the capacity for self-complementarityFigure 6-33 G:U base pairThe double helical structure of RNA resembles the A-form structure of DNA.The minor groove is wide a

23、nd shallow, but offers little sequence-specific information. The major groove is so narrow and deep that it is not very accessible to amino acid side chains from interacting proteins. Thus RNA structure is less well suited for sequence-specific interactions with proteinsStructure (2): RNA can fold u

24、p into complex tertiary structuresRNA STRUCTURE (3)RNA has enormous rotational freedom in the backbone of its non-base-paired regionsWhy?Interactions in the tertiary structure Unconventional base pairing, such as base triples, base-backbone interactions Proteins can assist the formation of tertiary structures by large RNA moleculeThe crystal structure of a 23S ribosme Function: Some RNAs are enzymesRNA STRUCTURE (4)Ribozymes are RNA molecules that adopt complex tertiary structure and serv