Biology Unit 1

Topic 2: Genes and Health

NUCLEIC ACID ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ Students will be assessed on their ability to: 1) Describe the basic structure of mononucleotides 2) Describe the structures of DNA and RNA 3) Describe how complementary base pairing and the hydrogen bonding between two complementary strands are involved in the formation of the DNA double helix ) Describe DNA replication !) "xplain how #eselson and $tahl%s classic experiment provided new data which supported the accepted theory of replication of DNA and refuted competing theories ~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~~ I T!UCTU!E "# NUCLE"TIDE Nucleic acid forms genetic materials of all living organisms Two types: DNA and RNA Long chain nucleotides 3 components: ! "-car#on sugar $ pentose - deo%yri#ose &in DNA! - ri#ose &in RNA! '! Nitrogenous #ase - purine $ ' rings $ adenine and guanine - pyrimidine $ ring $ thymine and cytosine and uracil - A( )( T and * in DNA - A( )( + and * in RNA ,hosphate group

3!

-ormation of nucleotide: - condensation of sugar . nitrogenous #ase . phosphate - two water molecules are removed -ormation of nucleic acid: - condensation #etween phosphate group of one with the sugar of the other $ phosphodieste$ %ond - %ac&%one of long polynucleotide chain $ alte$nating s'ga$ and phosphate - #ases pro/ecting sideways from sugars - se('ence o) %ases $ in)o$*ation which controls the organism0s development - "0 end $ phosphate group attached to car#on " - 30 end $ hydro%yl group on car#on 3

Biology Unit 1

Topic 2: Genes and Health

-ormation of mononucleotide t$'ct'$e o) DNA Nucleotide $ deo%yri#ose sugar( nitrogenous #ase &A( T( ) and *! and a phosphate group ' polynucleotide chains twisted around each other $ do'%le heli+ ' strands run in opposite direction - antipa$allel 1eld #y hyd$ogen %onds #etween nitrogenous #ases of opposite strands Co*ple*enta$y %ase pai$ing $ purine pairs with pyrimidine $ maintain constant width #etween ' sugar-phsophate #ac2#one *hargaff0s rule $ num#er of nucleotide with purine 3 num#er of nucleotide with pyrimidime 4ne complete turn of dou#le heli% 3 5 #ase pairs 3 distance of 367 nm

The importance of #ase pairing and hydrogen #onding: ! 8n DNA replication - se9uence of two strands are complementary - self-replication is possi#le $ each strand act as a template $ original DNA gives rise to ' copies $ identical #ase se9uence '! 8n DNA repair - alteration of removal of nucleotide from one strand - complementary strand act as template for repair - ensure integrity of DNA se9uence :aintain sta#ility of DNA molecule - purine pairs with pyrimidine $ constant width - hydrogen #onds holding each #ase - hydropho#ic interactions #etween stac2ed #ases - covalent phosphodiester #onds #etween ad/acent nucleotide - sta#ilise structure of dou#le heli%

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Biology Unit 1

Topic 2: Genes and Health

t$'ct'$e o) !NA ;ingle stranded ;ugar ri#ose instead of deo%yri#ose Nitrogenous #ase uracil instead of thymine ;ynthesised in the nucleus Three forms: messanger RNA( transfer RNA( ri#osomal RNA 8nvolved in protein synthesis II 3 $ "< of total RNA conveying message from nucleus to ri#osomes #ase se9uence complementary to DNA each triplet &codon! codes for one amino acid specify the order of amino acid in a polypeptide chain =5< of total RNA $ three different types genes situated at nucleolar organi>ers &secondary constriction! e%it nucleus and com#ine with proteins to form ri#osomes "< of total RNA a#out =5 ri#onucleotides long - clover shape $ folded #ac2 upon itself and held in shape #y hydrogen #onding #etween complementary #ase pairs at certain regions 30 end $ **A $ #inds to specific amino acid and "0 end $ end with ) anticodon $ specific triplet #ase se9uence complementary to a codon on mRNA at least '5 different tRNA molecules carry amino acid to ri#osome during protein synthesis

*!NA:

$!NA:

t!NA:

!E,LICATI"N "# DNA

Th$ee Alte$nati-e .odels o) DNA !eplication Three possi#le hypothesis can #e used to descri#e the process of DNA replication: ! *onservative replication $ would leave intact the original DNA molecule and generate a completely new molecule6 2) Dispersive replication would produce two DNA molecules with sections of #oth old and new DNA interspersed along each strand 3) ;emi-conservative replication would produce molecules with #oth old and new DNA( #ut each molecule would #e composed of one old strand and one new one

Biology Unit 1

Topic 2: Genes and Health

A classic e%periment #y :atthew :eselsohn and -ran2 ;tahl will allow you to differentiate #etween the three models6 Two isotopes of nitrogen( 7N and "N can #e distinguished on the #ases of their mass: 7 N is lighter than "N6 ?acteria are grown in a medium where their nitrogen source( used to synthesise nucleotides and eventually DNA is the ammonium ion &N1 7.! containing heavy "N6 After several generations all the DNA in these #acteria is heavy6 &' (here are the N atoms in the DNA) ;ome of the parental #acteria are now placed in a fresh medium containing light 7N ammonium ions and are allowed to divide once( producing generation 6 8f DNA replication is semi-conservative( all the generation hy#rid DNA with one heavy and one light strand6 &' "xplain why) #acteria are e%pected to have

;ome generation #acteria are now allowed to /ust divide once in 7N medium so producing generation ' #acteria6 8f DNA replication is semi-conservative( two types of #acterial DNA are now e%pected to #e present in generation '( i6e6 light and hy#rid6 &' "xplain why)

Biology Unit 1

Topic 2: Genes and Health

To confirm whether semi-conservative( replication is occurring( we now need to isolate and separate the DNA from generation and generation ' to see if the pattern of light and hy#rid DNA is as predicted6 This can #e done using the techni9ue of density gradient centrifugation which separates molecules on the #asis of their density6 After centrifugation( the tu#es are irradiated with +@ light of wavelength 'A5nm6 DNA a#sor#s this wavelength very strongly so the position of the DNA in the centrifuge tu#e shows up as #lac2 #and6 :eselsohn and ;tahl0s results are shown in figure #elow6 )eneration 5 B6coli with " N-la#elled DNA )eneration *ells replicate once on 7 N medium )eneration ' *ells replicate a second time on 7 N medium

DNA is e+t$acted and cent$i)'ged

&' &'

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(hy do these support the hypothesis that DNA replication is semi*conservative) +f the replication had been i) conservative ii) dispersive what pattern of DNA bands would be seen in the centrifuge tubes for generations 1 and 2) +f the bacteria had been left to divide for another two generations in the light ammonium medium, in what ratio would the hybrid and light DNA occur in the cells of generation )

Biology Unit 1

Topic 2: Genes and Health

e*i/conse$-ati-e !eplication o) DNA ?efore the start of DNA replication: - free nucleotides are manufactured in the cytoplasm - transported into nucleoplasm via nuclear pore ;tart of DNA replication: - #egins at origin of replication - #inding of en>ymes to the origin initiates replication - hydrogen #onds #etween complementary #ase pair are disrupted - separation of two parental strands - unwind ;ynthesis of new DNA strand: - ' parental strands act as templates - DNA polymerase catalyse synthesis - selects and align free deo%yri#onucleotides in se9uence complementary to the parental strand - in "0 to 30 direction - ad/acent deo%yri#onucleotides /oined #y phosphodiester #ond - ' daughter strands $ synthesised in opposite directions - continuous synthesis for one strand - lagging strand $ polymerase waited for new stretches on template to #e e%posed $ wor2 #ac2ward $ 42a>a2i fragment $ sugar-phosphate #ac2#one /oined #y ligase Bnd of replication: - complementary parental and daughter strands rewind into new heli% - se*i/conse$-ati-e $ newly formed dou#le heli% contains one polynucleotide chain of the original dou#le heli% and one newly synthesi>ed polynucleotide chain

Biology Unit 1

Topic 2: Genes and Health