(Foundation Block) Dr. Sumbul Fatma

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(Foundation Block) Dr. Sumbul Fatma Molecular biology (1) (Foundation Block) Dr. Sumbul Fatma

Learning outcomes To learn the central dogma of molecular biology. To have an understanding of the composition, types and structure of DNA and RNA. To have an idea about the organization of DNA in the chromosome and the role of histone proteins.

The central dogma of Molecular Biology A portion of DNA, called a gene, is transcribed into RNA. RNA is translated into proteins. Human genome contains about 35,000 genes

Nucleic acids Required for the storage and expression of genetic information. Two types: DNA (Deoxyribonucleic acid). RNA (Ribonucleic acid). Building blocks of nucleic acids are nuclueoside triphosphates (nucleotides).

Nucleotides Nucleotides are composed of: Nitrogenous base Pentose sugar Phosphate group

Nitrogenous base: Purines: Adenine (A) and Guanine (G) Pyrimidines: Cytosine (C), Thymine (T) and Uracil (U).

Sugar: pentose with 5 carbon ring: Ribose (with –OH at C2). Deoxyribose. Phosphate groups.

The sugar carbon numbers are primed (1’ 2’ 3’ etc The sugar carbon numbers are primed (1’ 2’ 3’ etc.), while the nitrogenous base atoms are unprimed. The nitrogenous base is bonded to C1’ of sugar. The PO4 group is bonded to C3’ or C5’ of sugar.

Function of nucleotides Polymers of nucleotides (as DNA or RNA) store and transfer genetic information. Free nucleotides and their derivatives perform various metabolic functions not related to genetic information. Other nucleotides: FAD, NAD, CoA.

Chemical structure of DNA & RNA The PO4 bridges the 3’ and 5’ positions of ribose sugar. The PO4 and sugar bonding is the backbone of DNA structure. The linkage between the nucleotides is called phosphodiester bond

The double helix DNA The structure of DNA was first determined by James Watson and Francis Crick in 1953. Commonly known as Watson-Crick structure.

Features of Watson-Crick DNA structure Two polynucleotide chains wind around a common axis to form a double helix. The two strands are anti-parallel (run in opposite direction). Each strand is a right-handed helix. The nitrogenous bases are in the center of the double helix and the sugar-phosphate chains are on the sides.

The surface of the double helix contains 2 grooves: the major and minor grooves. Each base is hydrogen bonded to a base in the opposite strand to form a base pair (A-T and G-C), known as complementary base pairing. The helix has 10 base pairs (bp) per turn.

Watson-Crick base pairs Adenine (A) Thymine (T) Guanine (G) Cytosine (C) In RNA, Thymine is replaced by Uracil (U)

Types of DNA structure A-DNA B-DNA* Z-DNA Direction Right-handed Left-handed Helix length Short Elongated More elongated Major groove Deep and narrow Wide Not real groove Minor groove Narrow Placement of bp Displaced away from the helical axis Centred over the helical axis Zig-zag pattern (nearly perpendicular to the helical axis) bp per turn 11 10 12 Conformation of deoxyribose C3 C2 G (C2) ; C (C3) * Watson-Crick model (B-DNA)

DNA supercoiling The chromosomes of many bacteria and viruses contain circular DNA which is supercoiled.

Melting temperature (MT) The temperature at which the double-stranded DNA is separated into two single strands. MT of DNA depends on nitrogenous base content (A-T and G-C). G-C has 3 hydrogen bonds  stronger than A-T.

RNA (Types and function) RNA is a single-stranded polymer of ribonucleotides. Types of RNA: mRNA (messenger RNA)  Function: Transcription process (from DNA to mRNA). tRNA (transfer RNA)  Function: Recognition and transferring. It recognizes amino acids’ codons and transfers the selected amino acids to the growing protein chain. rRNA (ribosomal RNA)  Function: Site of protein synthesis (factory).

Structure of a tRNA

How DNA is organized in a chromosome? The human genome contains 3.5 billion base pairs and more than 95% is non-coding or “junk” DNA. The DNA from single 23 human chromosomes have a length of 1 meter. How such large quantities of DNA are packed into a single cell? Each chromosome is a complex of a single linear DNA molecule and protein called chromatin. 50% of chromatin consists of proteins called histones.

Histones Five major types of histones: H1 H2A H2B H3 H4 Histones have positively charged amino acids (arginine and lysine). These proteins bind to negatively charged PO4 groups of DNA to stabilize the chromatin structure.

Nucleosomes Nucleosomes are particles consisting of DNA and histones connected by thin strands of naked DNA (like beads on a string; Sibhah in Arabic). Nucleosomes consist of the histone octamer (eight) and DNA (H2A)2(H2B)2(H3)2(H4)2 H1 binds to 2 complete helical turns of DNA.

Electron micrograph of chromatin showing nucleosomes

A nucleosome showing interaction of histones with the DNA

Chromatin filament with nucleosomes and naked DNA

Reference Lippincott’s Illustrated reviews: Biochemistry 4th edition – unit 2. Biochemistry by Voet and Voet 3rd edition. http://www.tulane.edu/~biochem/nolan/lectures/rna/bzcomp2.htm http://biowiki.ucdavis.edu/Genetics/Unit_I%3A_Genes,_Nucleic_Acids,_Genomes_ and_Chromosomes/Chapter_2._Structures_of_nucleic_acids/B-Form,_A-Form,_Z-Form_of_DNA