The adenine dinucleotide in combination with different vitamins form important Nicotinamide is a vitamin called Nicotinic acid compounds called eo. | (iaein) Flavin is. also a vitamin called enzymes. Three important eo. Niboflavin (Vitamin B2) cnéymes are NAD (Nicotinamide Adenine Dinucleotide), NADP (Nicotinamide denine Dinucleotide Phosphate) and FAD (Flavin Adenine Dinucleotide). These coenzymes can exist in two forms; a reduced and oxidized form. In the oxidized state they function as hydrogen acceptor. NAD and FAD are both active in the electron transport chain of respiration where they act as hydrogen carriers. i . _ Inthe process they are alternately reduced and oxidized. 4 : 0) P AD Adenine re Adenine \ Sone A : er Pp. Nicotinic Acid P Nicotinic Acid is See Structure of NAD ‘Structure of NADP Le Fig: 2.18 Some examples of co-enzymes 2.6.4 Polynucleotides (Poly-many) When many nucleotides are linked together they form a structure called polynucleotide. DNA and RNA molecules are polynucleotide chains (strand). ‘There are different types of RNA molecules, each type performs specific function under the instructions of DNA. Nucleic acids are of two types: 1. DNA: Deoxyribonucleic Acid — made of deoxyribonucleotides 2. RNA: Ribonucleic Acid - made of ribonucleotides Deoxyribonucleic Acid (DNA) In 1962 James Watson (b. 1928), Francis Crick (1916-2004), and Maurice Wilkins (1916-2004) jointly received the Nobel Prize in physiology or medicine for their 1953 determination of the structure of deoxyribonucleic acid (DNA). ‘According to this model the DNA molecule is double helix. The double helix can be visualized as spiral stair case wound around a central axis. Watson and Crick suggested that base pair always consists of purine pointing toward pyrimidines, Scanned with CamScannerReine i ee] keeping the molecule diameter at a constant 2nm. Th i i distance of 0.34 nm between them. nh sia a esi af \ ST ALCoW of DMA DNA contains pentose sugar as deoxyribose. It is formed of four different types of nucleotides, These nucleotides are named after the base present in them they are: Adenine deoxyribonucleotide, Guanine deoxyribonucleotide, Thymine deoxyribonucleotide and Cytosine deoxyribonucleotide. These four types of nucleotides are used as building blocks of DNA molecule. The nucleotides in the DNA molecule are bonded to one another in such a manner that the sugar of one nucleotide is linked to the phosphate group of the next one. In this way the nucleotides form a linear molecule called a strand in which the purines ogenous bases ‘ gp OO PUTT Guanip tains phucpher’e Qatil HaPO4 - 147 base paws make one Fig: 2.19 Structure of DNA nucleosome. backbone is made up of sugar alternating with the phosphate group. The bases are Projected to one side of the strand. The sequence of the nucleotide in one type of DNA is constant while it is different from other type of DNA molecule. DNA molecule consists of two strands. The two strands twist about one another in the form of a double helix. The two strands run in opposite direction to each other in the double helix. They are held together by hydrogen bonds between purine and Pytimidine bases. Thymine in one strand is always paired with ademnine in the opposite strand and guanine is always paired with cytosine. , M2: tinker oA joins ene Nudeescme Wh another ru gees - and _consigis Of 53 bGSE pUITS. So G NCE e _Aengnith tinker onA — QAO consysts of 2 mScanneres Continuation i fstrant Continuation Eero le Hydrogen bond There are two hydrogen bonds between adenine and thymine and three hydrogen bonds between guanine and cytosine. SS 2.6.5 Ribonucleic Acid Ribonucleic acid is also a polymer of the nucleotides, Ribonucleic acid called RNA contains pentose sugar as Ribose. It is also formed of four different types of nucleotides. These nucleotides are named after the base present in them they are: adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide, uracil ribonucleotide. The nucleotides in the RNA molecule Sto 3 direction are linked in the same manner as in the DNA molecule. RNA is a_ single polynucleotide strand. In RNA the base u racil occurs instead of the base thymine. oy ery adenine vam -, Cree an, Fig: 2.21 Chemical Structure of RNA Scanned with CamScanner‘There are three types of RNA molecules: mace (URNA) and ribosomal RNA (RNA Messenger RNA (mRNA), transfer RNA ger RNA (mRNA) . Transfer RNA (tRNA) Transfer RNA transfers the specifi Fi Fs © amino, ribosome for protein synthesis, abe sacid from the cytoplasm to the SS e, 2.6.6 DNA as Hereditary Material ; Chromosome is composed of DNA and Proteins. Biologists conducted experiments and Proved that DNA is the genetic material and i responsible for the transfer of genetic information from Parents to offspring, t oe In 1928 . Griffith conducted experiments using bacteria: that causing Pneumonia in* mice. He used two types Phage DNA in liquid et? 1 with radioactive ‘sulphur (35s) Batch 2: Phages grown fanradoacive phosphorus (32p) Fig. 23.6 Hershey and Chase Experiment Scanned with CamScannery 12: chapter 23 Chromosome and BNA’ a} cteria, Soon afler the infection 3 : arr gz05 » bacterial corp Ss wi sot ba 9 inte ntritugation technique, TI ere separa . pot eis ee ig an be medi contents and bacra, eae from media contents with the F355, In this analysis, vas found in the cells a 7 > clearly showed that during ‘nan bacterial Cells while 8g ei me i eae a rt ell while its 'S labelled en" “P labelled Dav + as found in the medium, These pe bacterial —_ Protein coat remained ._. OF bacteriophage was injected into outside the host. Based on these obse Sutside. Subsequently, many viral particles h ‘ vations, pe vis Protein, WAS FeSPOnSible for directing yne’ "MY Ad Chase claimed that the virue DNA, watle = eee 6 the production of new viruses (Fig. 23. TTR ef ee SSSR ribe the paradoxical nature of DNA, as a toot of geneticists and f say ope of rganism can be identified by examination of DATS Sequences creusies ofthat individual (sometimes Salted 8 DNA fingerprint). There is an extretnely small chance that another person has the same DNA profile ae One Sr Set of 13 regions. Examples of DNA used for ens 2 nl suspects whose DNA may match evidence lef at crime scenes (2) De i alread DNA REPLICATION ) making copy af SND he process of self-synthesis of DNA molecule is called DNA replication. This process occurs once in S-phase during the life cycle of a cell. The molecule of DNA which is replicated is called rent DNA, while the molecules, produced in this process are called daughter DNA. A parent DNA nolecule after replication gives rise two daughter DNA molecule, 133.1 Models of DNA Replication = : Pee aa How duplex DNA can replicate? Scientists started struggle to find the answer of this by the dscovery of DNA structure. Over all three different models to explain the replication process were seated ie., semi-conservative, conservative and dispersive model (Fig, 23.7). se the data to create a DNA profile Seni-conservative model . According to this model, the two parental DNA strands separate and each of these strands then Snes emplate forthe synthesis of anew DNA stand. The results 1wo DNA double eles, both, “Sihch consist of one parental and one new strand, — Conservative model . Parent PNA remains Same. ? This model states that after replication has occurred one daughter molecule contains both parental Strands, and the other daughter molecule contains DNA strands of all newly-synthesized material, Yinersive model : ; : 4 2 is model explains that during DNA replication, the parental DNA molecule is dispersed into its les, Scanned with CamScanner tide, then! these nucleotide combine together with new nucleotide to form two new daughter DNA. ul ¢by Conservative model Ss Soe SS se reatn ioe x 2. OS Fig. 23.7 Models of DNA replication ‘The three models of DNA replication were evaluated by Mathew M eer ae eer apm eal California Institute of technology in 1958. Later on, they were awarded Nobel Prize. In this experiment, it was concluded finally that the replication of DNA occurs according to semi-conservative model. They grew bacteria in a medium containing heavy isotope of nitrogen, ‘SN, which became incorporated into the bases of bacterial DNA. After several generations, the bacteria were shifted to three separate plates, which were already poured with medium containing '4N. Three DNA samples were taken from bacteria shifted from ‘SN medium to the “4N medium. Fist sample was obtained from first plate just after the transfer of culture; called sample at 0 minute, the second sample was taken from second plate after 20 minutes, called sample at 20 minutes, and third sample was taken from third plate after another 20 minute, called sample at 40 minute. In addition 0 these, a control sample was also taken from the bacteria which were grown separately in “N medium. ‘The DNA samples were dissolved in cesium chloride (CsCl) solution and then spun at a very hig? speed in an ultra-centrifuge for many hours. The cesium and chloride ions tend to be pushed bY ceiitrifugal force towards the bottom of the tube. Ultimately a gradient of Cs* and CI’ ions WS established in the tube. Molecules of DNA were settled down and formed sediments to the level of the appropriate densities in test tubes. DNA of control sample was appeared lightest as formed sediment at the top of test tube, while DNA of sample at 0 minute was appeared heaviest as it formed sediment at the bottom of test tube- DNA of sample at 20 minute formed sediment intermediate level to that of control sample and sample 0 minute whereas sample at 40 minute had two sediments, one at the top and other at intermediate ev"! ig. 23.9)- Scanned with CamScanner42: Chapter 23 Chromosome and DI oo en : Bact SA ‘Bacterla grown in Sennsrer eae sown gon Ny a se eer 3 mm Some 4. Sample at snmics (Sy Snboes SOL, Serasee ena } "| Suspended DNA. | [eects prey ‘Conttitugation both ; nas Ng hy bria 3 ‘ ; ys ee Om Fig. 23.9 Heselson-Stahl experiment Meselson-Stahl interpreted their results as follows: . |Tepot ¢DNA of control sample appeared lightest because it had nate ‘strands of MN, whereas DNA of sample at 0 minute Yared heaviest because it had both strand of N, but | * fitst round of replication each daughter duplex was a wit Possessing one’strand of MN and one of '*N, so it |Botom ot oe ie sine at intermediate level. When this hybrid [higher density) Generation *« replicated in second round of replication, it “'ibuted "SN strand to form another hybrid duplex and Fig. 23.8 Results of Meselson-Stahl experiment Scanned with CamScanner208 nds that is why this sample formed ty, they claimed that the DNA replication is sep My strar ‘SN strand to form a light duplex containing both "'N strat sediments. On the basis of above mentioned results, conservative (Fig. 23.8). 23.3. anism of DNA Replication es a : t here we are going to discu: Although DNA feplication is a continuous process but phases for the sake of convenience. ee aie phase is characterized by the formation of replication bubble and replication fork, which are formed at a particular site, called origin of replic: ion site. Kista specific Sequence of nucleotides along the length of DNA from where process of replication begins. In cukaryotic DNa, there may be more than one origin of replication sites but in prokaryotic DNA there is only one origin of replication Replication bubble is formed when DNA gyrase (topoisomerase) and DNA helicase enzymes work at-origin of replication. DNA gyrase opens the turns of DNA duplex so the DNA is converted from spiral ladder like form to straight ladder like form. At the same time DNA helicase breaks down the base pairs of DNA so the two strands gradually separate from each other and give a bubble like appearance at origin of replication. Afier the breakdown of base pairs, the single strands of DNA are prevented to pair up again by specific proteins called single stranded binding (SSB) proteins, Boh single strands of DNA will act as template strand in the next phase and direct the synthesis of daughter strands along themselves. Each side of replication bubble is now termed as replication fork. Extcusion / Polymerization phase _ Extension or polymerization is referred to the formation of daughter strands (leading or lagging strands) along the template strands. The daughter strands are actually synthesized by DNA polymerase enzyme but this enzyme cannot work unless some nucleotides are arranged on template. For this Purpose primase enzyme is involved fo arrange some nucleotides on template strands. Such sti fragments of few nucleotides are called primers. Each primer is short oli i RNA acts as start site for the activity of DNA polymerase, oligonucleotide strand of Afier the establishment of primers, synthesis of daught ymers 7 ter sti i enzyme. There ate three diferent forms ofthis enzymes "ss Deeits by the DNA pot a) DNA polymerase-I: Tepairing process of DNA damages during the life ™ t is the main enzy F template daring replication process, "St S¥MMesizes both daughter strands slots 4 se 4 Scanned with CamScannerSLUG LT. Pinpaeay Fig. 23.10 Fig. 23.10 DNA replication (@) Replication fork (b) Formation of Okazaki fragment Mechanism OF DNA polymerasectI aetivity This enzyme is a huge dimer molecule ic., ab units. DNA polymerase-III cannot initiates pecxisting 3-OH group, and, therefore, needs sids nucleotide at 3" end of primer so the din pabunit also possesses ability to remove wron proofreading. Both units of DN, work on one template it consists of two units that further consist of-several replication process, it can add a nucleotide onto only a @ primer to perform its polymerase activity. It always ection of replication becomes 5' to 3! end. One of its ig nucleotide if it is added mistakenly. This ability is called ‘A polymerase-III are interlinked by a small polypeptide chain, first unit and continuously synthesize a daughter strand towards replication fork, this ntinuously growing daughter strand is called leading strand, while the second unit work on other ‘mplate and synthesizes another daughter strand away from replication fork. As the two units are iterlinked so the second unit is allowed to polymerize daughter strand up to a specific length, then it ' 0 jump back (100 to 200 nucleotides in prokaryotes and 1000 to 2000 nucleotides in eukaryotes) to ‘new primer to perform polymerization again, Therefore, this daughter strand grows discontinuously ‘vay from the replication fork by forming short fragments interrupted by primers, called Okazaki fagments. This discontinuously growing strand is called Ingging strand, tion phase inaticy ce’ ig characterized by the replacement of primers by DNA nucleotides and Okazaki fagneats in lagging strand to form a continuous strand. 7 imers by DNA nucleotides is carried out by DNA polymerase-t that has Fiat potent of primers by “00 guts os exonuclease. Its attached tothe 3 end af Oke thai beside polymerst fe oo that it can extend while on the other hand it cleaves ide from 5 ary ee this way primers are removed and each Okazaki fragment is up eae a ee Ki fragment but they do not join together. ee = faaments is carried out by DNA ligase enzyme that finally constructs acces of Okezatt Carat fragments so continuous strand is formed (Fig. 23.10). lester bone vetween Scanned with CamScanner210 See process of semi conservative replication of DNA. = 7 a duri tion. = how DNA conserves one strand, during replicat | Describe how various sci DNA replication. cba —— c The Polymerase Chain Reaction (PCR), is the most revolutionary technique used in biotechnology which is based 5 vitro DNA replication, This isan extremely sensitive means of amplifying small quantities of ON in the detection of low level bacterial infections or rapid changes in transcription at the single cel tion of a specific individual's DNA in forensic science. It can also be used in DNA sequencing, disorders, site specific mutation of DNA, or cloning or sub cloning of cDNAS. =a “DNA is the genetic material which can replicate. The genetic material is in the form of specific sequences of nucleotides along the DNA strands. Now the question arises how does this information, determine the traits of an organism? How is its message translated by cells into a specific trait? The DNA inherited by an organism forms specific traits by directing synthesis of a protein that acts as catalyst and catalyses a specific chemical reaction of the cell. Thus a gene expresses itself in a protein or enzyme that controls the development of a specific character or function. We can say that proteins are link between the genotype and phenotype. Gene expression or protein synthesis includes transcription and translation. The idea that DNA makes protein via an intermediate RNA is known as the central dogma of molecular genetics. Information can only flow from DNA to protein and not-from proteins to DNA. la other words, changes in DNA may change the resulting protein, but changes in proteins cannot feedback and change the DNA. Transcription is the synthesis of RNA from DNA. It is the first step of gene expression. It oo in Go, Gi and Gz phases of cell cycle. Unlike DNA replication, it requires only one enzyme 10 completed i.e., RNA polymerase. However, it is a continuous process; for convenience we can divide! into three phases: initiation, elongation and termination. ‘ Promoter DNA Coding = 38 sequence cond ~10 sequence Fig. 23.11 Structure of promoter region Scanned with CamScannery ] 42: Chapter 23 Chromosome and DNA” rs . i e 241 joltatton phase qranscription begins with the binding of RNA Polymerase at omoter region. In prokaryotes, there are two bin , fated in promoter ic. TATAAT also called -10 sequenee and | TGACA also called -35 sequence, whereas in eukaryotes, TATA ‘ATA box) also called -25 sequence and CAAT (CAAT box) also aaled -70 sequence. Names of these sequences (-10, -35 or -25, - 70) refer to position that these sequences are located before the initiation site of structural region of the gene (Fig. 23.11), iding sites are RNA polymerase consists of four subunits: alpha, beta, beta’ _Fi8- 23.12 Structure of RNA and sigma; only the first three subunits are required for polymerase Leta) activity and are considered the core enzyme while the sigma factor is required for RNA polymerase to bind to the promoter. It is similar to the DNA polymerase in that it also adds nucleotides to the 3' end of the growing polypeptide chain but unlike DNA polymerase it does not require primer to perform polymerase activity. In prokaryotes, only one type of RNA polymerase is found while in eukaryotes, there three types of RNA polymerases, namely RNA polymerase-l, which synthesize rRNA, RNA olymerase-II, which synthesize mRNA, RNA polymerase-III which synthesize tRNA (Fig. 23.12). As the RNA polymerase binds to-the promoter, DNA duplex become unwind, base pairs are - broken down, and a bubble like structure, the transcription bubble is appeared. Elongation phase As the RNA polymerase binds to promoter, sigma factor is released and remaining core enzyme extends the polymerization of ribonucleoside triphosphates (rNTP). It does not require primer to initiate polymerization. One of the two strand of the gene acts as {emplate for transcription. This template strand is also called antisense because mRNA is complementary to this strand. The other strand of the gene is called coding or sense strand. In elongation phase, RNA polymerase keep on moving from 5' to. added tothe end of the RNA RNADNA ‘hybrid region . Fig. 23.13 Elongation phase of transcription Scanned with CamScannerao . Biology 12: Chapter 23 Chromosome shat bubble also moves along the DNA " directi i i ide it transcription ‘ . : 3' direction towards the terminator region, beside it tr ipl This event continues till the RNA leaving the growing RNA strand protruding from the bubble. polymerase reaches the terminator region of the gene (Fig. 23-13). Termination phase The sequence of terminator region of the gene stops . i . The part region consists of a series of GC base pairs followed by a series of AT po oe fs toe mRNA which is transcribed in this region, projects to form a loop like structure c2 Chal ie ‘ owed by a small tail of poly U nucleotides, The GC hairpin causes the RNA polymerase t0 S'OP Te SYHNeSis of RNA (Fig. 23.14). the synthesis of mRNA. The terminaty RNA Ser EEE polymerase complementary . sequence a se uuuuUU we ‘3° Hairpin loop (GC-rich) RNA Fig. 23.14 Termination of Transcription In prokaryotes, there is no delay between trans mn and translation as the mRNA emer from DNA it undergoes translation due to the absence of a definite nucleus. On the other hand ® eukaryotic cells the pre-mRNA (newly synthesized mRNA) has to be modified into mature functional mRNA because transcription occurs in nucleus while translation in cytoplasm so mRNA bi to. move from nucléus to cytoplasm and during this journey, enzymes like phosphatases and nucleas® may degrade it, Further, eukaryotic mRNA also contains some non-coding region called introns Wi are removed or spliced during this process. Post transcriptional modification is therefore involved " events, addition of a cap and tail to protect it from degradation and RNA splicing to remove non- © sequences (Fig. 23-15). . Scanned with CamScanner— Intron ra, [n° [ear Gad) [a oP pases aan CBP . e Splicing occurs: Cyloplaam Fig: 23.15 mRNA processing in eukaryotes ‘A cap is in the form of 7-methyl GTP, which is liked from its 5! to the 5' end of mRNA. A ification also takes place at the opposite end of the RNA transcript in the form of a small chain of 500 adenine nucleotides, called poly-A tail, which is attached to the 3" end of the mRNA. These two difications prevent the mRNA to be degraded by phosphatases and nucleases. The removal of introns and maturation of primary mRNA to secondary or functional mRNA is called RNA splicing, Splicing is catalysed by the spliceosomes which is a large RNA-protein complex. Later on the spliced exon fragments are joined together with the help of RNA ligase enzyme. Ithas been discussed earlier in this chapter that a gene is a specific segment of DNA that encodes the synthesis of a particular polypeptide (protein). Therefore, the order of amino acids in a polypeptide is according to the sequence of nucleotides in any part of a DNA. This relationship between amino acids sequence and nucleotide sequence is called genetie code. In other words the information in DNA for the Synthesis of protein is referred to as genetic code which is transcribed’ copied into mRNA. Scanned with CamScanner. i fee ee 214 Biology — ‘The mRNA transcript consists of a random sequence of only four kind of nitrogenous bases (A, ¢ C and U). Now the question is that kow can a code with four letter alphabet specif’ a protein which a any given point contains one renty di i ids? Clearly a single base cannot specif, my give i e venty different amino acids? ; annot specify y given point contains of twenty only four different amino acids could single amino acid, if one base specifies a single amino acid thet fc be “encoded for, and proteins cabiining only four different amino acids would re fet’. Nor is jt feasible for just two bases to specify a single amino acid since only 16 amino ee sae 3 eno for (4 X 4 = 16). But three bases are sufficient. With three bases a total of 4X4 = 64 combinations are possible; each is specific for a particular amino acid. Therefore a triplet of bases along the length of mRNA that specifies a particular amino acid is called codon. There are total 64 codons. Three of these codons act as stop codons (UGA, UAG, and UAA), one of these must be present at the end of mRNA that indicates that the message is over. Since, these three codons do not encode any amino acid, hence called nonsense codon, while all the other sequences that encode specific amino acids are called sense codons. One of these sense codons (AUG = methionine) also acts as start codon which must be present at the beginning of all the sequences that code for amino acid chains, Genetic code has following characteristics: 1) Some amino acids are only encoded by a single codon; while others are encoded by up to four codons and amino acids leucine and serine are encoded by six codons. This property is known as code degeneracy. 2 The genetic code is universal. It is the same in almost all the organisms. For example AGA specifies arginine in bacteria, in humans and all other organisms whose genetic’ code has been studied. Because of the universality of codon the genes can be transferred from one organism to another and be successfully transcribed and translated in their new host. ‘Second letter (base) First letter (base) (eseq) 393191 PUL Fig.23.16 Types of codons = Scanned with CamScannercay 12: Chapter 23 Chromosome and DNA fe sway of genetic code of mitochondtial DNA however, showed Wa BERG co ‘versal. For example: UGA codon i 5 ‘ Teversal. FOr exa codon is normally'a stop codon but in mitochondr yophan, Likewise, AUA was read as methionine instead of oleate sna ne temination of protein synthesis instead of arginine, - ae sat: 4 There is no punctuation between each codon. It means that there is no gap between two codons. y) The genetic code is non-over lapping. mRNA sequence AUGAGCGCA is not read as AUGIUGA/GAG ctc., it will be read as AUG/AGC/GCA. - Translation is second step of gene expression. In translation, messenger RNA (mRNA) produced ty transcription is decoded by the ribosome to produce a specific amino acid chain, or polypeptide, that will ater fold into an active protein. Although translation is a continuous process but for convenience we will discuss it in four phases: activation of amino acids, formation of initiation complex, elongation snd termination. — i Activation of amino acids [anime acto} c= 44 Translation ‘Activation of athino acids refers to the binding of seoveting ’ fee amino acids dispersed in cytoplasm to the 3 end of enzyme be particular RNA molecules, in this way a complex is + formed called aminocyl-tRNA complex. This binding is catalyzed by aminocyl-tRNA synthase (activation enzyme). Various amino acids that are to take part in polypeptide formation have been continuously activated ‘oughout the process of translation (Fig. 23.17). fon of initiation complex ess of translation actually begins with the Fig. 23.17 Activation of amino acids of initiation complex. It is formed by the o ‘ation of ribosomal subunits, mRNA and first aminocyl-tRNA complex. First a (RNA molecule ig a chemically modified methionine (called N-formyl methionine) binds to the smaller ribosomal it. This binding is controlled by ; ‘enzyme called initiation factor. At same time 5’ end of mRNA y lecule also binds to the smaller Sub quate d a wnit of ribosome with the help of = # 7 aso ‘ther initiation factor. Initiation “A plex is completed when, larger subunit of ribosome is also, placed smaller subunit (Fig. 23.18). Sees Fig. 23.18 Formation of initiation complex = Scanned with CamScanner21 CNPP 22. MeCN The region of smaller ribosomal subunit where first aminocyltRNA complex is attached is calleg P site (peptidyl site), here peptide bonds will be formed between successive amino acids during elongation phase. Nearby two other sites arc also established. A site (aminocyl site) where successive tRNAs bearing amino acids will be attached and (exit site) where empty RNAS Will leave ribosome during elongation phase. Polypeptide elongation In this phase ribosomal units move along mRNA, amino acids are brought by tRNAs, which are joined together to form a polypeptide chain. This is accomplished by three steps which are repeateg again and again throughout this phase. a) Whichever codon of mRNA is exposed at A site, its anticodon bearing aminocyl-RNA complex binds to it with the help of an enzyme, the elongation factor. b) Then an enzyme peptidyl transferase is emerged from P site. It removes the amino acid (may be a chain) from tRNA present on P site and binds it to the newly coming amino acid with the help of peptide bond. c) Then ribosomal sub-units slightly move along mRNA from 5' to 3’ direction so that a new codon is exposed at A site. This movement is called translocation. As a result, the empty tRNA is reached at E site to leave the ribosome, while the other tRNA bearing a chain of amino acid is shifted from A site to P site, and another codon is exposed to A sité. These three steps are repeated again and again until the stop codon is reached at A site (Fig. 23.19). Fig. 23.19 Chain elongation in translation process Scanned with CamScanner= Termination Elongation continues if this fast Nonsense codons do not bind to any process of translation ribosome and the ribos i __ PROKARYOTES hion until a chain-ten tRNA, but they and the polypeptide is relcas, somal subunits Separate from tI inating nonsense codon is exposed at A site. ite recognized by release factors that terminate the ed from the tRNA.The tRNA is released from the the mRNA. (Fig. 23.20), EUKARYOTES Structure of ribosomes 1. Ribosomes are smaller in size consist of a 50S large subunit and a 30S small subunit, which bind together to form 70S ribosome, 1. Ribosomes are larger in size consist of a 60S large subunit and a 40S small subunit, which bind together to form 80S ribosome. Site of synthesis 2, It occurs in cytosol. Initiation phase — 2. It occurs in cytosol and in rough endoplasmic reticulum. initiating amino acid is modified ie. N- ‘methionine. specific purine-rich sequence on the 5" side is jired to distinguish initiator AUG from internal . mRNA is poly cistronic i.e. it can serve as a iplate for the synthesis of several proteins. nly one type of initiation factor is used 3. The initiating amino acid is methioni modified. . 4. There is no need of a specific purine-tich sequence instead the AUG nearest the 5’ end ofm RNA is usually selected as the start site. 5. mRNA is mono cistronic ice. it can serve as a template for the synthesis of only one protein. 6. More than one type of initiation factors are used ine. It is not ars _SRUTT factors are used. thesized protein J.TWwo release Utilization of syn tination’ 77, Single release factors is used’ There are several possible ways of utilization of newly synthesized protein within or outside of “lls. The Golgi apparatus prepares the proteins fo yr use both inside and outside the cell.Proteins are ‘Wed everywhere in the cell, doing all of different complex functions, Some proteins are incorporated in enbranes while other are incorporated into mitocl hondria and the nucleus. Proteins act as enzymes. TOteins take part in contraction ¢.g., actin, myosin and tubulin. Structural proteins include collagen, Scanned with CamScanner