The Molecular Basis of Inheritance Flashcards
scientists believed that proteins made up genes/inherited material
- proteins are major component of all cells
- complex macromolecules in seemingly limitless variety + great specificity of function
- a lot known about structure, very little known about DNA
Griffith (1927)
- experiments with different strains of bacterium
- some strains virulent and causes pneumonia, some strains harmless
- discovered that bacteria have ability to transform harmless cells into virulent ones by transferring some genetic factor from one bacteria cell to the other
- bacetrial transfromation + transformation experiment
Avery, MacLeod, McCarty (1944)
- published findings that Griffith’s transformation factor is DNA
- DNA was agent that carried genetic characteristics
- provided direct experimental evidence that DNA, not protein, was the genetic material
Hershy and Chase (1952)
- experiments that had strong support to DNA theory
- tagged bacteriophages with radioactive isotopes (32P and 35S)
- proteins contain sulfur but not phosphorus (DNA vice versa) so 32P labeled DNA of phage viruses while 35S labeled protein coat of phage viruses
- when bacteria infected with phage viruses, 32P always entered bacterium while 35S remained outside
- proved that DNA from viral nucleus, not protein from viral coat, was infecting bacteria and producing thousands of progeny
Rosalind Franklin (1950-53)
carried out X-ray crystallography analysis of DNA that showed helix
Watson and Crick (1953)
- **proposed double helix structure of DNA **
- used data from other scientists to build their model
- 2 major pieces of data used were the biochemical analysis of DNA (Erwin Chargaff) and X-ray diffraction analysis of DNA (Rosalind Franklin)
Meselson and Stahl (1958)
- proved that DNA replicates in semiconservative fashion (as Francis Crick predicted)
- cultured bacteria in medium with heavy nitrogen, allowing bacteria to incorporate heavy nitrogen into DNA as they replicated/divided
- bacteria then transferred to medium with light nitrogen and replicate/divided once
- bacteria then spun in centrifuge and found to be midway in density between bacteria grown in heavy/light nitrogen
- new bacteria contained DNA consisting of 1 heavy strand and 1 light strand
Structure of DNA
-double helix like twisted ladder, consist of 2 strands running in opposite direction (5’ to 3’ and 3’ to 5’)
- polymer consisting of repeating units of nucleotides, these consist of** 5-carbon sugar (deoxyribose), a phosphate, and a nitrogen base**
- carbon atoms in deoxyribose numbered 1 to 5
- four nitrogenous bases (adenine, thymine, cytosine, guanine: adenine and quanine purines, thymine and cytosine pryimidines)
- nitrogenous bases paired by hydrogen bonds: adenine bonds by double hydrogen bond to thymine, cytosine bonds by triple hydrgoen bond to guanine
Structure of DNA
-double helix like twisted ladder, consist of 2 strands running in opposite direction (5’ to 3’ and 3’ to 5’)
- polymer consisting of repeating units of nucleotides, these consist of** 5-carbon sugar (deoxyribose), a phosphate, and a nitrogen base**
- carbon atoms in deoxyribose numbered 1 to 5
- four nitrogenous bases (adenine, thymine, cytosine, guanine: adenine and quanine purines, thymine and cytosine pryimidines)
- nitrogenous bases paired by hydrogen bonds: adenine bonds by double hydrogen bond to thymine, cytosine bonds by triple hydrgoen bond to guanine
structure of DNA
deoxyribonucleic acid
- double helix like twisted ladder, consist of 2 strands running in opposite direction (5’ to 3’ and 3’ to 5’)
- polymer consisting of repeating units of nucleotides, these consist of** 5-carbon sugar (deoxyribose), a phosphate, and a nitrogen base**
- carbon atoms in deoxyribose numbered 1 to 5
- four nitrogenous bases (adenine, thymine, cytosine, guanine: adenine and quanine purines, thymine and cytosine pryimidines)
- nitrogenous bases paired by hydrogen bonds: adenine bonds by double hydrogen bond to thymine, cytosine bonds by triple hydrgoen bond to guanine
- gets packed and unpacked in nucleus as needed
- eukryotic DNA combines with large amounts of histones (protein) from which it seperates briefly during replication
- DNA + histones = chromatin
- double helix of DNA wraps twice around core of histones, forming nucleosomes
structure of RNA
ribonucleic acid
- single-stranded helix with repeating nucleotides: A, C, G, U (uracil which replaces T)
DNA replication in eukaryotes
- double helix unzips, and each strand is a template for formation of new strand with complementary nucleotides
- replication begins at sites called origins of replication where 2 strands seperate to form replication bubbles
- 1000s of bubble seen along DNA molecule, and speeds up process of replication along giant DNA molecule with 6 billion nucleotides, bubbles expands as replication proceeds in both directions at once
- replication fork at ends of replication bubbles (Y-shaped region where new strands of DNA elongating), all bubbles eventually fuse
- enzyme DNA polymerase catalyzes antiparallel elongation of new DNA strands
- DNA polymerase builds new strand from 5’ to 3’ direction by moving along template strand and pushing replication fork ahead (50 nucleotides/second elongation in humans)
- DNA polymerase cannot initiate synthesis but can only add nucleotides to 3’ end of existing chain (consist of RNA and called RNA primer, enzyme primase makes primer by joining RNA nucleotides)
- DNA polymerase replicates 2 original strands of DNA differently, even though it builds new strands in 5’ to 3’ direction, 1 strand formed towards replication fork in unbroken/linear fashion (leading strand)
- lagging strand forms in direction away from replication fork in series of segments (Okazaki fragments) which are 100-200 nucleotides long + joined into continuous strand by enzyme DNA ligase
- other proteins + enzymes assist in replication: helicases is enzyme that untwist double helix at replication fork which seperates 2 parental strands
- single-stranded binding proteins is scaffolding, holding strands apart
- topoisomerases lessen tension on tightly wound helix by breaking, swiveling, and rejoing DNA strands
- DNA polymerases carry out mismatch repair (proofreading that corrects errors), damaged regions of DNA excised by DNA nuclease
- everytime DNA replicates, some nucleotides from ends of chromosomes are lost, so special nonsense nucleotide sequences at ends of chromosomes that repeat thousands of times
- protective ends called telomeres and are created and maintained by enzyme telomerase
- telomeres get shorter everytime, clock that counts cell divisions and causes cell to stop dividing as cell ages
a cell can replicate entire DNA in a few hours
DNA to protein
- triplet code in DNA transcribed into codon sequence in mRNA inside nucleus
- newly formed strand of RNA (pre-RNA) then processed or modified in nucleus
- codon sequence leaves nucleus and is translated into amino acid sequence (polypeptide) in cytoplasm at ribosome
3 RNA types involved in protein synthesis
3 RNA types directly involved in protein synthesis:
1. Messenger RNA (mRNA) in transcription: when sequence of DNA expressed, 1 strand is copied into mRNA according to base-pairing rules
2. Ribosomal RNA (rRNA) in translation: structural and makes up ribosome, which consists of 2 subunits (large + small) with 1 mRNA binding site and 3 tRNA binding sites (A, P, E)
3. Transfer RNA (tRNA) carries amino acid from cytoplasmic pool of amino acids to mRNA at ribosome: shaped like coverleaf and and has binding site for an amino acid at one end and another binding site for anticodon sequence that binds mRNA at the other
transcription
process by which information in DNA sequence copied into complementary RNA sequence
3 stages of transcription
initiation, elongation, termination
initiation
- begins when enzyme, RNA polymerase recognizes and binds to DNA at promoter region
- promoter tells RNA polymerase where to begin transcription and which two strands to transcribe
- colllection of proteins (transcription factors) recognize key area within promoter (the TATA box, named for its repeating thymine and adenine nucleotides) and mediate binding of RNA polymerase to DNA
- completed assembly of transcription factors and RNA polymerase bound to promoter called transcription initiation complex
- one RNA polymerase attached to promoter, DNA transcription of DNA template begins
elongation
- elongation of strand continues as RNA polymerase adds nucleotides to 3’ end of growing chain
- RNA polymerase pries two strands apart and attaches RNA nucleotides according to base pairing rules
- stretch of DNA transcribed into mRNA molecule is called a transcription unit
- each unit consists of triplets of bases called codons that code for specific amino acids
- single gene can be transcribed into mRNA simultaneously by several molecules of RNA polymerase following each other
- has mechanisms for proofreading during transcription
termination
final stage, elongation continues for short distance after RNA polymerase transcribes termination sequence (AAUAAA), mRNA is cut free from DNA template
RNA processing
before pre-RNA strand shipped out of nucleus to ribosome, it is altered by enzymes:
- ** 5’ cap** (with a modified guanine nucleotide) added to 5’ end, helps RNA strand bind to ribosome during translation
- poly (A) tail (a string of adenine nucleotides) added to 3’ end, protects RNA strand from degradation by hyrolytic enzymes, and facilitates release of mRNA from nucleus into cytoplasm
- noncoding regions of mRNA called introns/intervening sequences spliced by SNPs (small nuclear ribonucleoproteins) within splicesomes, allows only exons (expressed sequences) to leave nucleus
- mRNA that leaves nucleus is a lot shorter than original transcription unit
alternative splicing
different RNA molecules produced from same primary transcript, depending on which RNA segments are exons vs. introns
regulatory proteins specific to cell type controls intron-exon choices by binding to regulatory sequences within primary transcript
translation of mRNA
- process by which codons of mRNA sequence are changed into amino acid sequence
- amino acids present in cytoplasm carried by tRNA molecules to codons of mRNA strand at ribosome according to baes-pairing rules
- one end of tRNA w/ specific amino acid, and other end w/ anticodon (nucleotide triplet)
- amino acid joined to correct tRNA by enzyme aminoacyl-tRNA synthetase
- 20 different aminoacyl-tRNA synthetase (1 for each amino acid), 64 codons (61 coding for amino acids, AUG codes for amino acid methione and is also a start codon, UAA UGA and UAG are stop codons and terminate translation)
- some tRNA molecules with anticodons to recognize 2+ different codons because pairing rules for third base of codon are not as strict as first 2 bases (relaxation known as wobble)
tRNA is used repeatedly unlike mRNA, energy provided by GTP (molecule similar to ATP)
initiation
in translation
- begins when mRNA attached to subunit of ribosome
- first codon is always AUG and must be positioned correctly for transcription of amino acid sequence to begin
elongation
in translation
- continues as tRNA brings amino acids to ribosome and polypeptide chain is formed
- a mRNA molecule translated simultaneously by several ribosomes in clusters (polyribosomes)
termination
in translation
- termination of mRNA strand complete when ribosome reaches one of three stop codons
- release factor breaks bond between tRNA and last amino acid of polypeptide chain
- poly peptide freed from ribosome, and mRNA is broken down
The Genetic Code
- 64 possible combinations of the 4 bases
- there are redundancies in the code, but no ambiguity
- code is universal and unifies all life, indicating that code originated early in evolution
gene mutation
- permanent changes in genetic material which occur spontaneously and randomly
- can be caused by mutagenic agents (chemicals/radiation)
- mutations in somatic cells disrupts normal cell function
- mutations in gametes transmitted to offspring and changes gene pool of population
- mutations are the raw material for natural selection
- some regions of DNA more vulnerable to mutation (A + T more breakages because of double bond rather than triple)