Friday, March 4, 2011

THE BIOCHEMICAL REACTIONS

  • DNA replication begins with the "unzipping" of the parent molecule as the hydrogen bonds between the are broken.
  • Once exposed, the sequence of bases on each of the separated strands serves as a template to guide the insertion of a complementary set of bases on the strand being synthesized.
  • The new strands are assembled from 
  • Each incoming nucleotide is covalently linked to the "free" 3' carbon atom on the pentose (figure) as
  • the second and third phosphates are removed together as a molecule of (PPi).
  • The nucleotides are assembled in the order that complements the order of bases on the strand serving as the template.
  • Thus each C on the template guides the insertion of a G on the new strand, each G a C, and so on.
  • When the process is complete, two DNA molecules have been formed identical to each other and to the parent molecule.

THE ENZYMES

  • A portion of the double helix is unwound by a
  • A molecule of aDNA polymerasebinds to one strand of the DNA and begins moving along it in the 3' to 5' direction, using it as a template for assembling aleading strand of nucleotides and reforming a double helix. In eukaryotes, this molecule is called DNA polymerase delta (δ).
  • Because DNA synthesis can only occur 5' to 3', a molecule of a second type of DNA polymerase (epsilon, ε, in eukaryotes) binds to the other template strand as the double helix opens. This molecule must synthesize discontinuous segments of polynucleotides (calledfragments). Another enzyme, then stitches these together into the 
DNA Replication is Semiconservative
When the replication process is complete, two DNA molecules — identical to each other and identical to the original — have been produced. Each strand of the original molecule has
  • remained intact as it served as the template for the synthesis of
  • a complementary strand.
This mode of replication is described as semi-conservative: one-half of each new molecule of DNA is old; one-half new. Watson and Crick had suggested that this was the way the DNA would turn out to be replicated. Proof of the model came from the experiments of Meselson and Stahl. [

Bacteria

The single molecule of DNA that is the genome contains 4.7 x 106nucleotide pairs. DNA replication begins at a single, fixed location in this molecule, the proceeds at about 1000 nucleotides per second, and thus is done in no more than 40 minutes. And thanks to the precision of the process (which includes a "proof-reading" function), the job is done with only about one incorrect nucleotide for every 109 nucleotides inserted. In other words, more often than not, the E. coli genome (4.7 x 106) is copied without error! 

Eukaryotes

The average human chromosome contains 150 x 106 nucleotide pairs which are copied at about 50 base pairs per second. The process would take a month (rather than the hour it actually does) but for the fact that there are many places on the eukaryotic chromosome where replication can begin. Replication begins at some replication origins earlier in S phase than at others, but the process is completed for all by the end of S phase. As replication nears completion, "bubbles" of newly replicated DNA meet and fuse, finally forming two new molecules. 
With their multiple origins, how does the eukaryotic cell know which origins have been already replicated and which still await replication? 
An observation: When a cell in G2 of theis fused with a cell in S phase, the DNA of the G2 nucleus does not begin replicating again even though replication is proceeding normally in the S-phase nucleus. Not until mitosis is completed, can freshly-synthesized DNA be replicated again. 
Two control mechanisms have been identified — one positive and onenegative. This redundancy probably reflects the crucial importance of precise replication to the integrity of the genome. 


In order to be replicated, each origin of replication must be bound by:
  • an OriginRecognition Complex of proteins (ORC). These remain on the DNA throughout the process.
  • Accessory proteins called licensing factors. These accumulate in the nucleus during G1 of the cell cycle. They include:
    • Cdc-6 and Cdt-1, which bind to the ORC and are essential for coating the DNA with
    • MCM proteins. Only DNA coated with MCM proteins (there are 6 of them) can be replicated.
Once replication begins in S phase, 
  • Cdc-6 and Cdt-1 leave the ORCs (the latter by and destruction in 
  • The MCM proteins leave in front of the advancing replication fork.


G2 nuclei also contain at least one protein — called geminin — that prevents assembly of MCM proteins on freshly-synthesized DNA (probably by blocking the actions of Cdt1). 
As the cell completes mitosis, geminin is degraded so the DNA of the two daughter cells will be able to respond to licensing factors and be able to replicate their DNA at the next S phase.

DNA Translation


Translation occurs in the cytoplasm where the ribosomes are located. Ribosomes are made of a small and large subunit which surround the mRNA.
Transfer RNA (tRNA) molecules are 75 - 95 nucleotides long and have four arms and three loops. True to its name, tRNA transfers amino acids to the site of the growing protein chain (polypeptide). Each tRNA molecule (in red below) recognises a specific, three base-pair mRNA code or codon (the DNA form of a codon is called a triplet and the sequence on the tRNA is called an anticodon). Since there are three bases and four possible nucleotides, there can be up to 64 (4x4x4) possible tRNA molecules. Three of these tRNA molecules recognise "stop" or termination codons which have been named amber (UAG), opal (UGA) and ochre (UAA).
The codon indicates which amino acid is to be added and the amino acid is attached to the tRNA molecule at the acceptor arm. As we can see from the table below, most amino acids are represented by more than one codon. This means that the expected protein can still be synthesised, even when a degree of mutation occurs in the DNA or mRNA.
There are 20 essential amino acids, however they can be combined in any order, just like the four nucleotides. This permits the production of the many different proteins which let organisms grow and function.
Name
1-Letter Nickname
Triplet
3-Letter Nickname
Glycine
G
GGT,GGC,GGA,GGG
Gly
Alanine
A
GCT,GCC,GCA,GCG
Ala
Valine
V
GTT,GTC,GTA,GTG
Val
Leucine
L
TTG,TTA,CTT,CTC,CTA,CTG
Leu
Isoleucine
I
ATT,ATC,ATA
Ileu
Serine
S
TCT,TCC,TCA,TCG,AGT,AGC
Ser
Threonine
T
ACT,ACC,ACA,ACG
Thr
Cysteine
C
TGT,TGC
Cys
Methionine
M
ATG
Met
Glutamic Acid
E
GAA,GAG
Glu
Aspartic Acid
D
GAT,GAC,AAT,AAC
Asp
Lysine
K
AAA,AAG
Lys
Arginine
R
CGT,CGC,CGA,CGG,AGA,AGG
Arg
Asparagine
N
AAT,AAC
Asn
Glutamine
Q
GAA,GAG
Gln
Phenylalanine
F
TTT,TTC
Phe
Tyrosine
Y
TAT, TAC
Tyr
Tryptophan
W
TGG
Trp
Unknown
X
 
xxx
Proline
P
CCT,CCC,CCA,CCG
Pro
Terminator
*
TAA,TAG,TGA
End


INITIATION
When the large ribosmal subunit,small ribosomal subunit, mRNA and the tRNA carrying a methionine come together in the cytoplasm, the ribosome becomes active and the synthesis of a polypeptide, or "translation", is initiated. The AUG codon binds at theprotein binding site (P) of the ribosome and AUG is always the first codon of an mRNA.





The next complementary tRNA will bind at theattachment binding site(A) of the ribosome. The adjacent amino acids are then joined by a peptide bond via a peptidaseenzyme. Thus the polypeptide chain begins to grow and as it does, it is passed to the next tRNA currently occupying the A site.






ELONGATION
The ribosome then moves 1 codon down the mRNA in a 5' to 3' direction. This is achieved by atranslocase enzyme. As the process of ribosome translocation continues, the "old" tRNA is released to bind another amino acid and go in search of a new codon. The binding of a new aminoacid is mediated by an enzyme called amino-acyl-tRNA synthase






TERMINATION
The process continues along the mRNA until a stop codon is reached. While there is no tRNA for a stop codon, there is an enzyme calledrelease factor which cleaves the polypeptide chain resulting in a new protein.





Finally, the entire complex is disrupted, the ribosome separates and the mRNA is released to be used again or degraded. Translation occurs at multiple sites along an mRNA so that many ribosomes can be seen by electron microscopy bound to a single mRNA strand with many polypeptide chains forming from each.

Introduction to DNA Structure

COMPONENTS OF DNA

DNA is a polymer. The monomer units of DNA are nucleotides, and the polymer is known as a "polynucleotide." Each nucleotide consists of a 5-carbon sugar (deoxyribose), a nitrogen containing base attached to the sugar, and a phosphate group. There are four different types of nucleotides found in DNA, differing only in the nitrogenous base. The four nucleotides are given one letter abbreviations as shorthand for the four bases.
  • A is for adenine
  • G is for guanine
  • C is for cytosine
  • T is for thymine

PURINE BASES

Adenine and guanine are purines. Purines are the larger of the two types of bases found in DNA. Structures are shown below:

Structure of A and G

The 9 atoms that make up the fused rings (5 carbon, 4 nitrogen) are numbered 1-9. All ring atoms lie in the same plane.

PYRIMIDINE BASES

Cytosine and thymine are pyrimidines. The 6 stoms (4 carbon, 2 nitrogen) are numbered 1-6. Like purines, all pyrimidine ring atoms lie in the same plane.

Structure of C and T

DEOXYRIBOSE SUGAR

The deoxyribose sugar of the DNA backbone has 5 carbons and 3 oxygens. The carbon atoms are numbered 1', 2', 3', 4', and 5' to distinguish from the numbering of the atoms of the purine and pyrmidine rings. The hydroxyl groups on the 5'- and 3'- carbons link to the phosphate groups to form the DNA backbone. Deoxyribose lacks an hydroxyl group at the 2'-position when compared to ribose, the sugar component of RNA.

Structure of deoxyribose

NUCLEOSIDES

A nucleoside is one of the four DNA bases covalently attached to the C1' position of a sugar. The sugar in deoxynucleosides is 2'-deoxyribose. The sugar in ribonucleosides is ribose. Nucleosides differ from nucleotides in that they lack phosphate groups. The four different nucleosides of DNA are deoxyadenosine (dA), deoxyguanosine (dG), deoxycytosine (dC), and (deoxy)thymidine (dT, or T).

Structure of dA

In dA and dG, there is an "N-glycoside" bond between the sugar C1' and N9 of the purine.

NUCLEOTIDES

A nucleotide is a nucleoside with one or more phosphate groups covalently attached to the 3'- and/or 5'-hydroxyl group(s).

DNA BACKBONE

The DNA backbone is a polymer with an alternating sugar-phosphate sequence. The deoxyribose sugars are joined at both the 3'-hydroxyl and 5'-hydroxyl groups to phosphate groups in ester links, also known as "phosphodiester" bonds.

Example of DNA Backbone: 5'-d(CGAAT):

Features of the 5'-d(CGAAT) structure:

  • Alternating backbone of deoxyribose and phosphodiester groups
  • Chain has a direction (known as polarity), 5'- to 3'- from top to bottom
  • Oxygens (red atoms) of phosphates are polar and negatively charged
  • A, G, C, and T bases can extend away from chain, and stack atop each other
  • Bases are hydrophobic

DNA DOUBLE HELIX

DNA is a normally double stranded macromolecule. Two polynucleotide chains, held together by weak thermodynamic forces, form a DNA molecule.

Structure of DNA Double Helix

FEATURES OF THE DNA DOUBLE HELIX

  • Two DNA strands form a helical spiral, winding around a helix axis in a right-handed spiral
  • The two polynucleotide chains run in opposite directions
  • The sugar-phosphate backbones of the two DNA strands wind around the helix axis like the railing of a sprial staircase
  • The bases of the individual nucleotides are on the inside of the helix, stacked on top of each other like the steps of a spiral staircase.

BASE PAIRS

Within the DNA double helix, A forms 2 hydrogen bonds with T on the opposite strand, and G forms 3 hyrdorgen bonds with C on the opposite strand.

Example of dA-dT base pair as found within DNA double helix

Example of dG-dC base pair as found within DNA double helix

  • dA-dT and dG-dC base pairs are the same length, and occupy the same space within a DNA double helix. Therefore the DNA molecule has a uniform diameter.
  • dA-dT and dG-dC base pairs can occur in any order within DNA molecules

DNA HELIX AXIS

The helix axis is most apparent from a view directly down the axis. The sugar-phosphate backbone is on the outside of the helix where the polar phosphate groups (red and yellow atoms) can interact with the polar environment. The nitrogen (blue atoms) containing bases are inside, stacking perpendicular to the helix axis.

View down the helix axis


What is DNA?


Deoxyribonucleic acid (/diˌɒksiˌraɪbɵ.njuːˌkleɪ.ɨk ˈæsɪd/  ( listen)), or DNA, is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms, with the exception of some viruses. The main role of DNA molecules is the long-term storage of information. DNA is often compared to a set of blueprints, like a recipe or a code, since it contains the instructions needed to construct other components of cells, such as proteins and RNA molecules. The DNA segments that carry this genetic information are called genes, but other DNA sequences have structural purposes, or are involved in regulating the use of this genetic information.



DNA consists of two long polymers of simple units called nucleotides, with backbones made of sugars and phosphate groups joined by ester bonds. These two strands run in opposite directions to each other and are therefore anti-parallel. Attached to each sugar is one of four types of molecules called bases. It is the sequence of these four bases along the backbone that encodes information. This information is read using the genetic code, which specifies the sequence of the amino acids within proteins. The code is read by copying stretches of DNA into the related nucleic acid RNA, in a process called transcription.

Within cells, DNA is organized into long structures called chromosomes. These chromosomes are duplicated before cells divide, in a process called DNA replication. Eukaryotic organisms (animals, plants, fungi, and protists) store most of their DNA inside the cell nucleus and some of their DNA in organelles, such as mitochondria or chloroplasts. In contrast, prokaryotes (bacteria and archaea) store their DNA only in the cytoplasm. Within the chromosomes, chromatin proteins such as histones compact and organize DNA. These compact structures guide the interactions between DNA and other proteins, helping control which parts of the DNA are transcribed

A DNA Spray Keeps Burglars at Bay




That gadget was  a canister loaded with a harmless solution containing synthetic DNA. If a criminal attempts to burgle a premises fitted with the device, an employee can hit a panic button that alerts police to a crime in progress and simultaneously shoots out a fine mist covering everyone in the room, including the robber. And as each batch of the spray -- which glows blue under ultraviolet light -- has a unique DNA signature, police can connect the robber to the scene of the crime.
A company-provided picture shows how the DNA spray fingers a burglar under ultraviolet light.


Criminally minded readers might now be thinking, "Well, if I robbed a shop, I'd just scrub myself clean when I got home." But as Andrew Knights, managing director of SelectaDNA, explains, the solution isn't so easy to remove. "It will come off within a number of hand washes," he told AOL News. "But if you run through a spray it'll accumulate on the inside of your nostrils and ears and under the fingernails; areas that are difficult to get off." And, he notes, if a criminal doesn't have an ultraviolet light, he won't know where the liquid is lurking.

Hundreds of sprays have been deployed at retailers and banks across the U.K.,and New Zealand; Knights' company, part of Britain's Selectamark security group, is also in talks with U.S. companies.

Knights says the sprays can reduce crime levels, but he admits the unique DNA evidence they offer has yet to be used in a prosecution. 

That's not a sign of failure, though. If a suspect is scanned with a UV light at a police station (almost everyone arrested in the U.K now undergoes this procedure, no matter what crime the person is suspected of) and starts to glow, he says, "They'll generally plead guilty. The criminal knows it's better to make a plea bargain, rather than annoy the police even further by forcing them to go through the DNA testing."

That's exactly what happened when an 18-year-old burglar from the town of Rawtenstall -- some 20 miles east of Preston in northwestern England -- was hauled in for questioning last month. When he walked under a UV light at the station, his arms started to shine, explains Police Constable Phil Buck, a crime-prevention coordinator in the northern English county of Lancashire. The young offender picked up the glow when he broke into a garden center whose roof had been smeared in another crime-fighting substance: SelectaDNA Gel. "He held up his brightly glowing hands and confessed," says Buck, adding that the teen admitted breaking into the gardening store three times.

However, the main aim of the SelectaDNA spray isn't to capture criminals but to scare them away. "Retailers are investing in this technology because they want to move the crime on somewhere else," Knights says. "They are just out to protect their property and staff." That's why every business that uses a SelectaDNA spray also prominently displays a bright yellow sign in their window showing a stick man with a bag of swag being hit by the mist. "Warning," the sign reads. "SelectaDNA spray installed here."

What is RNA?

Ribonucleic acid (RNA) is a biologically important type of molecule that consists of a long chain of nucleotide units. Each nucleotide consists of a nitrogenous base, a ribose sugar, and a phosphate.
RNA comes in a variety of different shapes. Double-stranded DNA is a staircase-like molecule.

RNA is very similar to  but differs in a few important structural details: in the cell, RNA is usually single-stranded, while  is usually double-stranded; RNA nucleotides contain ribose while  contains deoxyribose (a type of ribose that lacks one oxygen atom); and RNA has the base uracil rather than that is present in 
Ribonucleic acid (RNA) has the bases adenine (A), cytosine (C), guanine (G), and uracil (U).
Ribonucleic acid (RNA) has the bases adenine (A), cytosine (C), guanine (G), and uracil (U). Image Credit: National Institute of General Medical Sciences
RNA is transcribed from by enzymes called RNA polymerases and is generally further processed by other enzymes. RNA is central to protein synthesis. Here, a type of RNA called messenger RNA carries information from to structures called ribosomes. These ribosomes are made from proteins and ribosomal RNAs, which come together to form a molecular machine that can read messenger RNAs and translate the information they carry into proteins. There are many RNAs with other roles – in particular regulating which are expressed, but also as the genomes of most
RNA and  are both nucleic acids, but differ in three main ways. First, unlikewhich is double-stranded, RNA is a single-stranded molecule in most of its biological roles and has a much shorter chain of nucleotides. Second, while contains ''deoxyribose'', RNA contains ''ribose'' (there is no hydroxyl group attached to the pentose ring in the 2' position in These hydroxyl groups make RNA less stable than  because it is more prone to hydrolysis. Third, the complementary base to adenine is not as it is in  but rather uracil, which is an unmethylated form of  For instance, determination of the structure of the ribosome—an enzyme that catalyzes peptide bond formation—revealed that its active site is composed entirely of RNA.

What is DNA?

Deoxyribonucleic acid (DNA) is a nucleic acid that contains the genetic instructions used in the development and functioning of all known living organisms and some The main role of DNA molecules is the long-term storage of information. DNA is often compared to a set of blueprints or a recipe, or a code, since it contains the instructions needed to construct other components of cells, such as proteins and molecules. The DNA segments that carry this genetic information are called  but other DNA sequences have structural purposes, or are involved in regulating the use of this genetic information.
Chemically, DNA consists of two long polymers of simple units called nucleotides, with backbones made of sugars and phosphate groups joined by ester bonds. These two strands run in opposite directions to each other and are therefore anti-parallel. Attached to each sugar is one of four types of molecules called bases. It is the sequence of these four bases along the backbone that encodes information. This information is read using the genetic code, which specifies the sequence of the amino acids within proteins. The code is read by copying stretches of DNA into the related nucleic acid in a process called transcription.
DNA Structure
Within cells, DNA is organized into long structures called  Theseare duplicated before cells divide, in a process called DNA replication. Eukaryotic organisms (animals, plants, fungi, and protists) store most of their DNA inside the cell nucleus and some of their DNA in organelles, such as mitochondria or chloroplasts. In contrast, prokaryotes (bacteria and archaea) store their DNA only in the cytoplasm. Within the chromatin proteins such as histones compact and organize DNA. These compact structures guide the interactions between DNA and other proteins, helping control which parts of the DNA are transcribed.


DNA Preservation



To determine the effect of environmental factors on the preservation of DNA, archeological teeth of approximately similar age but greatly differing site milieu were examined for DNA content. The complex relational system of locational milieu of the samples was reduced to its essential and, at the same time, easily measurable factors. These are temperature, humidity, pH value, the geochemical properties of the soil, the amount of postmortal organic substances and the general degree of microbial infestation in the respective soil. The relative DNA content in the samples was established by determining the rate of successful polymerase chain reaction (PCR) amplifications. Differences in quantity and quality of the results are attributed to the respective prevailing environmental factor or to the respective storage conditions. Dryness, low temperature and absence of microorganisms favors the preservation of DNA. The bioapatite of bones and teeth, like the DNA, are preserved under neutral or slightly alkaline conditions. Brief storage at room temperature does not affect the amount of amplifiable DNA but does affect the reproducibility of the results. Long storage outside a lab freezer reduces the amount and the reproducibility of DNA amplifications in ancient specimens.

Structure And Diagrams Of Dna Replication

Structure And Diagrams Of Dna Replication

Dna Replication Structure And Diagrams


Dna Replication Structure And Diagrams 1


Dna Replication Structure And Diagrams 2


Dna Replication Structure And Diagrams 3


Dna Replication Structure And Diagrams 4


Dna Replication Structure And Diagrams 5


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