Erwin Chargaff determined that: 3-D structure of DNA: amount of adenine roughly equals the amount of thymine DNA is helical amount of cytosine roughly equals the amount of guanine diameter = 2nm makes a complete turn of the helix every 3.4 nm
1 10/22/2024
WHAT THE DIAMETER TELLS US: JAMES WATSON AND FRANCIS CRICK
proposed double helix structure
2 sugar-phosphate backbones nitrogenous bases toward interior of molecule complementary bases pair and are held together with hydrogen bonds
DNA STRUCTURE COMPLEMENTARY BASE-PAIRING
the two strands of
nucleotides are antiparallel to each other one is oriented 5’ to 3’ other is oriented 3’ to 5’
DNA STRUCTURE DNA REPLICATION – WATSON & CRICK “Now our model for deoxyribonucleic acid is, in effect, a pair of templates, each of which is complementary to the other. We imagine that prior to duplication the hydrogen bonds are broken, and the two chains unwind and separate. Each chain then acts as a template for the formation on to itself of a new companion chain, so that eventually we shall have two pairs of chains, where we only had one before. Moreover, the sequence of the pairs of bases will have been duplicated exactly.”
2 10/22/2024
DNA REPLICATION, THE BASIC CONCEPT DNA REPLICATION – CONSERVATIVE MODEL HYPOTHESIS
DNA REPLICATION – SEMI-CONSERVATIVE MODEL HYPOTHESIS DNA REPLICATION: DISPERSIVE MODEL
DNA REPLICATION – ORIGINS OF REPLICATION
replication fork = a Y-shaped region where the parental strands of DNA are being unwound helicases untwist the double helix & separate the two parental strands single-strand binding proteins keep the unpaired DNA strands from re-pairing topoisomerase helps relieve this strain created by the untwisting of the double helix
3 10/22/2024
SYNTHESIZING A NEW DNA STRAND
unwound sections of parental DNA strands are now available to serve as
templates enzymes that synthesize DNA require a primer primer = a complementary RNA chain, usually 5-10 nucleotides long
DNA polymerase adds DNA nucleotides to the 3’ end of a preexisting
chain rate of elongation is about 50 nucleotides per second in human cells
ANTIPARALLEL ELONGATION REPLICATION OF THE LEADING STRAND
DNA polymerases can add nucleotides only to the free 3’ end, never to the 5’ end i.e., a DNA strand can elongate only in the 5’ → 3’ direction one DNA strand, the leading strand, can be synthesized continuously the other DNA strand must work away from the replication fork this strand, the lagging strand, is synthesized discontinuously, as a series of segments called Okazaki fragments the two strands are produced simultaneously
DNA polymerases proofread each nucleotide against its template as soon
as it is covalently bonded to the growing strand. incorrectly paired nucleotide are removed mismatched nucleotides sometimes evade proofreading mismatch repair = other enzymes remove and replace incorrectly paired nucleotides
REPLICATING THE ENDS OF DNA MOLECULES
the usual replication machinery cannot complete the 5’ ends of
daughter DNA strands because there is no 3’ end of a preexisting polynucleotide for DNA polymerase to add onto repeated rounds of replication produce shorter and shorter DNA molecules with uneven (“staggered”) ends
TELOMERES typically consist of multiple repetitions of a short nucleotide sequence in humans the six-nucleotide sequence TTAGGG is repeated between 100 and 1000 times telomeres are how cells protect chromosome ends T-loop or “knot” at the very end of the telomer keeps the chromosome ends from sticking together each time a cell divides, the telomeres become slightly shorter (instead of genes become shorter or lost) eventually, they become so short that the cell can no longer divide successfully, and the cell dies in certain cell types that divide a lot, an enzyme called "telomerase" adds those repeats back so the telomere doesn't get too short
6 10/22/2024
TELOMERASE ACTIVITY & TUMORS
Normal shortening of telomeres may protect organisms from cancer by limiting the number of divisions that somatic cells can undergo. Cells from large tumors often have unusually short telomeres, as we would expect for cells that have undergone many cell divisions. Telomerase activity is abnormally high in cancerous somatic cells, suggesting that its ability to stabilize telomere length may allow these cancer cells to persist. Many cancer cells do seem capable of unlimited cell division, as do immortal strains of cultured cells
