UNIT IV: MOLECULAR BIOLOGY Module 10 (v): Protein synthesis: Transcription, Translation and Genetic code 10.1.

. Introduction: Chemically, proteins are polymers of amino acids. The amino acid molecules are linked together in a linear fashion forming long polypeptide chains. Protein synthesis includes arrangement of a definite number of amino acids in a definite sequential order. This sequence of the amino acids chain is determined by the sequence of nucleotides in the DNA. 10.2 What is Transcription? Transcription is the process in which RNA (ribonucleic acid) is synthesized from DNA. The word gene can be defined as the functional unit of the DNA that can be transcribed. Thus the genetic information stored in the DNA is expressed through RNA. One of the two strands of DNA serves as a template to produce working RNA molecules. This template is called non- coding or sense strand. The DNA strand which does not participate in the transcription is known as coding strand or anti sense strand. 10.3. Requirements for the transcription: i. DNA ii. RNA polymerase iii. NTPs iv. Promoter v. Rho factor Transcription in prokaryotes: RNA polymerase enzyme can synthesize all types of RNAs in prokaryotes. The overall transcription involves 3 stages: initiation, elongation, termination. Initiation: There will be a specific region called promoter, where the RNA polymerase binds to the DNA. After this binding, RNA polymerase unwinds the DNA strand for about 17 bps (base pairs) to form a transcriptional bubble.

Elongation: RNA is synthesized from 5 end to 3 end, antiparallel to the DNA template because of the action of the enzyme RNA polymerase. This enzyme utilizes the ribonucleotide triphosphates for the formation of RNA. The base pair sequence in the RNA is complementary to the template DNA strand and identical to that of coding strand except that RNA contains U instead of T. Termination: A specific protein called rho factor binds to the growing RNA to terminate the transcription. Rho factor also dissociates the RNA polymerase from DNA.

Transcription of DNA

10.4. Translation: Translation is needed to produce the protein molecule from RNA. Translation is the process by which the genetic code carried by messenger RNA (mRNA) directs the production of proteins from amino acids. 10.7. Requirements for translation: i. mRNA ii. ribisomes iii. tRNA iv. amino acids v. ATP and GTP vi. Initiation factors and Elongation factors Activation of amino acids: Amino acids are get activated and attached to the tRNAs before translation. Each tRNA is specific for each amino acid.

10.8. Procedure: Protein synthesis occurs in ribisomes. mRNA is read in 5 3 direction and the protein synthesis proceeds from N terminal to C terminal. Translation is divided in to 3 stages: Initiation, elongation, termination. Initiation: Initiation factors IF1 and IF3 bind to the free 30S subunit of ribosome. Then the mRNA attached to the 30S subunit. In the next step methionine t RNA binds with the 30S sub unit in the presence of GTP. After this step, 50S subunit attached to the 30S subunit to form complete 70S ribosome. GTP provides necessary energy. Elongation: Ribosomes elongate the polypeptide chain by a sequential addition of amino acids. Nearly 40 amino acids per second are added to the growing chain. Elongation factors EF- Tu, EF-Ts helps in elongation. 50S subunit of the ribosome contains three sites: P site A site and E site. At beginning tRNA occupies the P site. The subsequent tRNA, which decodes the next codons occupies the A site. Formation of peptide bond between amino acids: Because of the action of the enzyme peptidlytransferase, peptide bond forms between the first amino acid at P site and second amino acid at A site. Then the ribosome moves from 5 end to 3 end (moves to next codon) on the mRNA. This process is called translocation. The second tRNA moves from A site to P site. The first tRNA will go out. Now the third tRNA occupies the empty A site. This elongation process repeats again and again till it reaches the end signal.

Termination:

UAA, UAG, UGA are the stop codons and do not have any specific tRNAs to bind with them. If the stop codon occupies the A site, release factors binds to the codon and helps to release the newly synthesized protein through E site. 10.9. Check points: 1. Transcription is the process in which RNA (ribonucleic acid) is synthesized from DNA. 2. The genetic information stored in the DNA is expressed through RNA 3. DNA strand which participate in the transcription is known as non-coding strand, and the other one called coding strand. 4. RNA polymerase binds to the DNA at promoter region during transcription. 5. The base pair sequence in the RNA is complementary to the template DNA strand and identical to that of coding strand except that RNA contains U instead of T. 6. Rho factor helps in the termination of transcription. 7. The genetic code is the set of rules by which information encoded in genetic material is translated into proteins by living cells. 8. Triplet codon in a nucleic acid sequence usually specifies a single amino acid. 9. Four bases produce 64 different combinations of triplet codons. 10.Translation is the process by which the genetic code carried by messenger RNA (mRNA) directs the production of proteins from amino acids. 11.mRNA and tRNA binds to the ribosome. 12.50S subunit of the ribosome contains two sites: P site and A site 13.Peptidlytransferase enzyme responsible for the formation of peptide bond. 14.Ribosome moves from 5 end to 3 end on the mRNA. 15.UAA, UAG, UGA are the stop codons. 10.10. Questions: Short answer questions: 1. What is genetic code? 2. What are the Characters of genetic code? 3. Describe the elongation step in translation. 4. What are the steps in transcription? Briefly discuss them. Long answer questions:

1. Write a detailed essay on translation. 2. Discuss the transcription. Glossary Codon: It is the three nucleotide sequence Translation: Translation is the process in which sequence of nucleotides in mRNA is translated into the sequence of amino acids of a polypeptide chain. A Codon (Code word): Is three nucleotide sequence in mRNA which codes for a particular amino acid; whereas the genetic code is the sequence of nitrogenous bases in mRNA molecule, which encloses the information for the synthesis of protein molecule. Punctuation: The imitator and terminator codons are known as signals and this phenomenon is known as punctuation. P site: Peptidyl site or donor sites. A site: Amino acyl or acceptor site. E site: Exit site. MCQs 1. Gene can be defines as a. Part of DNA b. Part of RNA c. Functional part of DNA d. Protein Answer: C 2. DNA strand which participate in the transcription is a. Anti sense strand b. Sense strand c. Non coding strand d. Both B and C Answer: D 3. Rho factor helps in a. Termination of translation b. Initiation of transcription c. Elongation of translation

4.

5.

6.

7.

8.

9.

d. Termination of transcription Answer: D RNA polymerase responsible for a. Synthesis of new RNA b. Synthesis of new DNA c. Synthesis of new protein d. Synthesis of new gene Answer: A What are the stop codons? a. UAA, UAG, AUG b. UAA, UAG, UGA c. UAA, AUG, UGA d. UAA, UAG, UGG Answer: B AUG codes for a. Leucine b. Methionine c. It is a stop codon d. Phenul alanine Answer: B Activation of amino acids occurs a. Before transcription b. After translation c. During translation d. Before translation Answer: D During translation a. mRNA moves on ribosome b. ribosome moves on mRNA c. both mRNA and ribosome moves on each other d. they doesnt move Answer: B The first tRNA occupies which site in 50S subunit of ribosome?

P site A site E site T site Answer: A 10. How many combinations of triplet codons are available? a. 61 b. 64 c. 20 d. 4 Answer: B

a. b. c. d.