CDMB Overview
The Central Dogma of Molecular Biology
DNA REPLICATION
• Before a cell duplicates and is divided into new daughter cells through
either mitosis or meiosis, biomolecules and organelles must be
copied to be distributed among the cells.
• The process of DNA duplication is called DNA replication. Replication
follows several steps that involve multiple proteins called replication
enzymes and RNA. In eukaryotic cells, such as animal cells and plant
cells, DNA replication occurs in the S phase of interphase during the
cell cycle.
• Cells must replicate their DNA before they can divide. This ensures
that each daughter cell gets a copy of the genome, and therefore,
successful inheritance of genetic traits.
• DNA replication is a process that is essential to all cells.
DNA replication would not occur without enzymes that catalyze various steps
in the process. Enzymes that participate in the eukaryotic DNA replication
process include:
• DNA helicase - unwinds and separates double stranded DNA as it moves
along the DNA. It forms the replication fork by breaking hydrogen bonds
between nucleotide pairs in DNA.
• DNA primase - a type of RNA polymerase that generates RNA primers.
Primers are short RNA molecules that act as templates for the starting
point of DNA replication.
• DNA polymerases - synthesize new DNA molecules by adding nucleotides
to leading and lagging DNA strands.
• Topoisomerase or DNA Gyrase - unwinds and rewinds DNA strands to
prevent the DNA from becoming tangled or supercoiled.
• Exonucleases - group of enzymes that remove nucleotide bases from the
end of a DNA chain.
• DNA ligase - joins DNA fragments together by forming phosphodiester
bonds between nucleotides.
Initiation
Elongation
Termination
DNA TRANSCRIPTION
• DNA transcription is a process that involves transcribing genetic
information from DNA to RNA. The transcribed DNA message, or RNA
transcript, is used to produce proteins.
• DNA is housed within the nucleus of our cells. It controls cellular activity by
coding for the production of proteins. The information in DNA is not
directly converted into proteins, but must first be copied into RNA. This
ensures that the information contained within the DNA does not become
tainted.
• Transcription describes the process by which the genetic information
contained within DNA is re-written into messenger RNA (mRNA) by RNA
polymerase. This mRNA then exits the nucleus, where it provides the basis
for the translation of DNA. By controlling the production of mRNA in the
nucleus, the cell regulates the rate of gene expression.
Initiation
Elongation
Termination
• Elongation will continue until the RNA polymerase encounters a stop
sequence. At this point, transcription stops and the RNA polymerase
releases the DNA template.
• RNA polymerase moves along the DNA until it reaches a terminator
sequence. At that point, RNA polymerase releases the mRNA polymer
and detaches from the DNA.
• Modified from Ca
Pre-translational mRNA processing
• The mRNA which has been transcribed up to this point is referred to as pre-mRNA.
Processing must occur to convert this into mature mRNA. The processing includes:
5′ Capping
• Capping describes the addition of a methylated guanine cap to the 5′ end of mRNA. Its
presence is vital for the recognition of the molecule by ribosomes, and to protect the
immature molecule from degredation by RNAases.
Polyadenylation
• Polyadenylation describes the addition of a poly(A) tail to the 3′ end of mRNA. The poly(A)
tail consists of multiple molecules of adenosine monophosphate. This stabilises RNA,
which is necessary as RNA is much more unstable than DNA.
Splicing
• Splicing allows one genetic sequence to code for different proteins. This conserves genetic
material. It involves:
• Introns (non-coding sequences) are removed via spliceosome excision
• Exons (coding sequence) are then joined together by ligation
• It is sequence dependent and occurs within the transcript
• This allows many proteins to be made from a single pre-mRNA.
TRANSLATION
• Translation is a process by which the genetic code contained within a messenger
RNA (mRNA) molecule is decoded to produce a specific sequence of amino acids
in a polypeptide chain.
• It occurs in the cytoplasm following transcription and, like transcription, has three
stages: initiation, elongation and termination. In this article we will look at the
components and stages of DNA translation.
• The key components required for translation are mRNA, ribosomes, and transfer
RNA (tRNA).
• During translation, mRNA nucleotide bases are read as codons of three bases.
Each ‘codon’ codes for a particular amino acid. Every tRNA molecule possesses an
anticodon that is complementary to the mRNA codon, and at the opposite end
lies the attached amino acid. tRNA molecules are therefore responsible for
bringing amino acids to the ribosome in the correct order ready for polypeptide
assembly.
• It is important to know
that a single amino acid
may be coded for by more
than one codon. There are
also specific codons that
signal the start and the
end of translation.
• Aminoacyl-tRNA
synthetases are enzymes
that link amino acids to
their corresponding tRNA
molecules. The resulting
complex is charged and is
referred to as an
aminoacyl-tRNA.
Initiation
For translation to begin, the start
codon 5’AUG must be recognised.
This is a codon specific to the amino
acid methionine, which is nearly
always the first amino acid in a
polypeptide chain.
At the 5’ cap of mRNA, the small
40s subunit of the ribosome binds.
Subsequently, the larger 60s subunit
binds to complete the initiation
complex. The next step (elongation)
can now commence.
Elongation
• The ribosome has two tRNA binding sites;
the P site which holds the peptide chain
and the A site which accepts the tRNA.
• While Methionine-tRNA occupies the P site,
the aminoacyl-tRNA that is complementary
to the next codon binds to the A site, using
energy yielded from the hydrolysis of GTP.
• Methionine moves from the P site to the A
site to bond to new amino acid there, and
so the growth of the peptide has begun.
The tRNA molecule in the P site no longer
has an attached amino acid, and so leaves
the ribosome.
• The ribosome then translocates along the
mRNA molecule to the next codon, again
using energy yielded from the hydrolysis of
GTP. Now, the growing peptide lies at the P
site and the A site is open for the binding of
the next aminoacyl-tRNA, and the cycle
continues. The polypeptide chain is built up
in the direction from the N terminal
(methionine) to the C terminal (the final
amino acid).
Termination
References
• Reece, Jane B., and Neil A. Campbell. Campbell Biology. Benjamin Cummings, 2011.
• 'Donnell M, Langston L, and Stillman B. Principles and concepts of DNA replication in bacteria, archaea, and eukarya. Cold
Spring Harb Perspect Biol 2013; 5(7). [PMID: 23818497]
• Remus D, Beuron F, Tolun G, Griffith JD, Morris EP, and Diffley JFX. Concerted loading of Mcm2-7 double hexamers around
DNA during DNA replication origin licensing. Cell 2009; 139(4):719-30. [PMID: 19896182]
• Langston LD, Zhang D, Yurieva O, Georgescu RE, Finkelstein J, Yao NY, Indiani C, and O'Donnell ME. CMG helicase and DNA
polymerase ε form a functional 15-subunit holoenzyme for eukaryotic leading-strand DNA replication. Proc. Natl. Acad. Sci.
U.S.A. 2014; 111(43):15390-5. [PMID: 25313033]
• Sheu Y, and Stillman B. The Dbf4-Cdc7 kinase promotes S phase by alleviating an inhibitory activity in Mcm4. Nature 2010;
463(7277):113-7. [PMID: 20054399]
• Zou L, and Stillman B. Formation of a preinitiation complex by S-phase cyclin CDK-dependent loading of Cdc45p onto
chromatin. Science 1998; 280(5363):593-6. [PMID: 9554851]
• Hook SS, Lin JJ, and Dutta A. Mechanisms to control rereplication and implications for cancer. Curr. Opin. Cell Biol. 2007;
19(6):663-71. [PMID: 18053699]
• Vijayraghavan S, and Schwacha A. The eukaryotic Mcm2-7 replicative helicase. Subcell. Biochem. 2012; 62:113-34. [PMID:
22918583]
• Stillman B. DNA polymerases at the replication fork in eukaryotes. Mol. Cell 2008; 30(3):259-60. [PMID: 18471969]
• You Z, De Falco M, Kamada K, Pisani FM, and Masai H. The mini-chromosome maintenance (Mcm) complexes interact with
DNA polymerase α-primase and stimulate its ability to synthesize RNA primers. PLoS ONE 2013; 8(8):e72408. [PMID:
23977294]
References
• Plosky BS. More division of labor at the eukaryotic replication fork. Mol. Cell 2014; 56(4):467-8. [PMID: 25459878]
• Balakrishnan L, and Bambara RA. Flap endonuclease 1. Annu. Rev. Biochem. 2013; 82:119-38. [PMID: 23451868]
• Balakrishnan L, and Bambara RA. Okazaki fragment metabolism. Cold Spring Harb Perspect Biol 2013; 5(2). [PMID: 23378587]
• Singleton MR, Dillingham MS, and Wigley DB. Structure and mechanism of helicases and nucleic acid translocases. Annu. Rev.
Biochem. 2007; 76:23-50. [PMID: 17506634]
• Kegel A, Betts-Lindroos H, Kanno T, Jeppsson K, Ström L, Katou Y, Itoh T, Shirahige K, and Sjögren C. Chromosome length influences
replication-induced topological stress. Nature 2011; 471(7338):392-6. [PMID: 21368764]
• Bermejo R, Kumar A, and Foiani M. Preserving the genome by regulating chromatin association with the nuclear envelope. Trends
Cell Biol. 2012; 22(9):465-73. [PMID: 22771046]
• Kumar A, Mazzanti M, Mistrik M, Kosar M, Beznoussenko GV, Mironov AA, Garrè M, Parazzoli D, Shivashankar GV, Scita G, Bartek J,
and Foiani M. ATR mediates a checkpoint at the nuclear envelope in response to mechanical stress. Cell 2014; 158(3):633-46.
[PMID: 25083873]
• Shivashankar GV. Mechanosignaling to the cell nucleus and gene regulation. Annu Rev Biophys 2011; 40:361-78. [PMID: 21391812]
Online Links
DNA Translation - Initiation - Elongation - Termination – TeachMePhysiology
Translation of mRNA to Protein: Initiation, Elongation & Termination Steps - Video & Lesson Transcript | Study.com
Stages of transcription: initiation, elongation & termination (article) | Khan Academy
A Central Dogma Molecular Biology Glossary (miami.edu)
