The CENTRAL DOGMA of MOLECULAR BIOLOGY

The Central Dogma of Molecular Biology

formulated by Francis Crick states that the flow of genetic information is DNA to RNA to protein

The Central Dogma of Molecular Biology

forms the backbone of Molecular Biology represented by three major stages: DNA replication, transcription, and translation

DNA REPLICATION
the process of copying a double-stranded DNA molecule both strands serve as templates for the copying of the opposite strands

Models of DNA Replication

The two parental DNA strands are back together after replication has occurred. That is, one daughter molecule contains both parental DNA strands, and the other daughter molecule contains DNA strands of all newly-synthesized material.

Models of DNA Replication

The two parental DNA strands separate and each of those strands then serves as a template for the synthesis of a new DNA strand. The result is two DNA double helices, both of which consist of one parental and one new strand.

Models of DNA Replication

The parental double helix is broken into double-stranded DNA segments that, as for the Conservative Model, act as templates for the synthesis of new double helix molecules. The segments then reassemble into complete DNA double helices, each with parental and progeny DNA segments interspersed.

Models of DNA Replication

The Meselson and Stahl Experiment proved that the semiconservative model was the correct one.

Requirements for DNA replication

1. Master copy of DNA the parental DNA
2. Deoxyribonucleoside triphosphates (dATP, dGTP, dTTP, dCTP) serve as substrates 3. Helix-unwinding protein (HUP) to unwind the parental strands in order to create two template strands 4. Helix-destabilizing protein (HDP) prevent the separated strands from reannealing

Requirements for DNA replication (cont.)

5. Helix-relaxing protein (DNA gyrase) release the tension of supercoiled twist created in unwinding the parental DNA without rotation
6. DNA polymerase catalyzes the synthesis of the daughter DNA; requires a 3-OH end as a primer for the addition of the next nucleotide 7. RNA polymerase initiates the synthesis of RNA primer strands

Requirements for DNA replication (cont.)

8. 5 3 and 3 5 exonucleases for cleaving the RNA primer from the elongation 9. DNA ligase joins the nicks and gaps by catalyzing the formation of phosphodiester bonds

Biosynthesis of DNA
DNA polymerase catalyzes the synthesis of new DNA in the 5' to 3' direction using a DNA template strand.

The substrates for DNA polymerase are not the nucleotides but related molecules called nucleoside triphosphates.

Biosynthesis of DNA (cont.)

Nucleoside triphosphates have three phosphate groups instead of the one phosphate characteristic of nucleotides.

Biosynthesis of DNA (cont.)

In DNA biosynthesis, a nucleoside triphosphate base pairs with the base on the DNA template strand, and a phosphodiester bond is formed between the growing DNA chain and the new nucleotide.

Biosynthesis of DNA (cont.)

DNA polymerase both ensures that the correct base pair is formed and catalyzes the formation of the phosphodiester bond.

Steps of DNA replication

Step 1:

A. The DNA is already partially unwound to form a replication fork. B. On the bottom template strand, primase synthesizes a short RNA primer in the 5' to 3' direction.

Step 1 (cont.)

C. Primase leaves, and DNA polymerase adds DNA nucleotides to the RNA primer in the 5' to 3' direction. This new DNA is called the leading strand because it is being made in the same direction as the movement of the replication fork.

Step 2

A. On the top template strand, primase synthesizes a short RNA primer in the 5' to 3' direction.

Step 2 (cont.)

B. Primase leaves, and DNA polymerase adds DNA nucleotides to the RNA primer in the 5' to 3' direction. This new DNA is called the lagging strand because it is being made in the direction opposite to the movement of the replication fork. The segment produced is also called an Okazaki fragment.

Step 3

A. The DNA unwinds some more and the leading strand is extended by DNA polymerase adding more DNA nucleotides. Thus, the leading strand is synthesized continuously.

Step 4

A. On the top template strand, a new RNA primer is synthesized by primase near the replication fork and is DNA is added to it by DNA polymerase. This produces the second Okazaki fragment. Thus, the lagging strand is synthesized discontinously.

Step 4 (cont.)

B. DNA ligase joins the two Okazaki fragments to produce a continuous chain.

Step 4 (cont.)

C. The process repeats as the DNA continues to unwind. Because one new DNA strand is synthesized continuously and the other is synthesized discontinuously, this model is called the semidiscontinuous model for DNA synthesis

Step 5

A. A different type of DNA polymerase removes the RNA primer and replaces it with DNA. DNA ligase joins the two Okazaki fragments to produce a continuous chain. B. The process repeats as the DNA continues to unwind.

Summary: DNA replication

Summary: DNA replication

TRANSCRIPTION
the synthesis of an RNA copy (mRNA) of a segment of DNA

RNA is synthesized by the enzyme RNA polymerase

In prokaryotic cell

transcription and translation are coupled; translation begins while the mRNA is still being synthesized.

In prokaryotic cell

because there is no nucleus to separate the processes of transcription and translation, when bacterial genes are transcribed, their transcripts can immediately be translated.

In eukaryotic cell
transcription occurs in the nucleus, and translation occurs in the cytoplasm

In eukaryotic cell
transcription and translation are spatially and temporally separated in eukaryotic cells; transcription occurs in the nucleus to produce a premRNA molecule

In eukaryotic cell
the pre-mRNA is typically processed to produce the mature mRNA, which exits the nucleus and is translated in the cytoplasm.

Stages of transcription

TRANSCRIPTION
the process by which non-coding sequences of base pairs (introns) are subtracted from the coding sequences (exons) of a gene in order to transcribe DNA into messenger RNA (mRNA) in chromosomes, DNA acts as a template for the synthesis of RNA; in most mammalian cells, only 1% of the DNA sequence is copied into a functional RNA (mRNA)

TRANSCRIPTION (cont.)
only one part of the DNA is transcribed to produce nuclear RNA, and only a minor portion of the nuclear RNA survives the RNA processing steps one of the most important stages in RNA processing is RNA splicing in many genes, the DNA sequence coding for proteins, or "exons", may be interrupted by stretches of non-coding DNA, called "introns". In the cell

TRANSCRIPTION (cont.)
in many genes, the DNA sequence coding for proteins, or exons, may be interrupted by stretches of non-coding DNA, called introns in the cell nucleus, the DNA that includes all the exons and introns of the gene is first transcribed into a complementary RNA copy called nuclear RNA (nRNA)

TRANSCRIPTION (cont.)
introns are removed from nRNA by a process called RNA splicing the edited sequence is called messenger RNA (mRNA) the mRNA leaves the nucleus and travels to the cytoplasm, where it encounters cellular bodies called ribosomes to continue the translation process

TRANSCRIPTION (cont.)

TRANSLATION
the messenger RNA is decoded to produce a specific polypeptide according to the rules specified by the genetic code preceded by transcription

TRANSLATION (cont.)
also in 3 phases: initiation, elongation and termination (all describing the growth of the amino acid chain, or polypeptide that is the product of translation)

TRANSLATION (cont.) Initiation

the components of the translational apparatus come together with a molecule of mRNA a tRNA carrying the 1st amino acid (AA1) binds to the start codon

TRANSLATION (cont.)

Elongation
amino acids are conveyed to the mRNA by tRNAs and are added one by one, to a growing polypeptide chain

TRANSLATION (cont.) Termination

a stop codon in the mRNA is recognized by a protein release factor the translational apparatus comes apart, releasing a completed polypeptide