Endonuclease V-DNA Repair Enzyme
Endonuclease V also known as EndoV is a highly conserved DNA repair enzyme. According to the research done in past, this enzyme was first discovered in E-coli and then later homologs of this enzyme were also found in prokaryotes, humans, and eukaryotes. This biochemical analysis of this protein is still not well understood by the scientist; however, Thermotoga maritime (Tma) which is a thermophillic bacterium is used as a great model to explain its enzymatic properties and structural properties.
EnodoV is encoded by nfi gene and the main function of this DNA protein is to repair DNA bases that have been damaged due to the exposure to the ultra-voilet radiation. These damaged DNA bases are known as deaminated
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According to a journal cripts.org, EndoV has a crystal appearance and metal-binding with site attached to its surface along with hypoxanthine-binding pocket. Apart from these, it was discovered that its structure is similar RNase-H and it also showed some RNA-cleaving properties. Besides this, it has many more insertion on its surface for Cysteine dimers such as Cys227 and Cys228. These two cysteines attached to its surface play an import part in the catalytic function of EndoV. Studies have compared the bacterial and human EndonucleaseV and found that bacterial EndoV shows high cleavage efficiency on nucleic acid substrates, whereas human EndoV has shown more efficiency towards single stranded RNA and is most active towards DNA with …show more content…
And this property of Endo V allows other proteins to complete the downstream repair process. So this study signifies that acts as a strong DNA repair protein. Also the activity of EndoV is not complete without a metal factor of Mg2+ that is responsible for the catalysis of this enzyme. Also the metal factor upon binding with DNA it actually shows 3’ and 5’ non-specific exonuclease activity. This was demonstrated using a 5’ and 3’ labeled inosine containing DNA and H2124D mutant, where the actual 3’ exonuclease activity was
Figure 2 shows the results of the electrophoresis. Lanes 5 and 7 indicate the fragments obtained when the plasmids are digested with both restriction enzymes, indicating the approximate fragment size for the hlyA gene, the pK184 plasmid and the pBluescript plasmid. This is useful for identifying the recombinant DNA needed for this experiment
The endo product forms almost exclusively because of the activation barrier for endo being much lower than for exo. This means that the endo form is formed faster. When reactions proceed via the endo for the reaction is under kinetic control. Under kinetic control the adduct is more sterically congested, thus thermodynamically less stable. The endo form has a lower activation energy, however, the EXO form has a more stable product.
In 1994, a stable in Hendra, a suburb of Brisbane, Queensland, Australia broke out with an unknown respiratory disease that resulted in thirteen horses and one horse trainer severely ill, resulting in death. [7, 8] This disease was isolated by scientists and later classified as the Hendra virus. The Hendra virus (HeV), previously unknown, is now classified under the family Paramyxoviridae, genus Henipavirus along with its sister viruses the Nipah Virus and Cedar virus.[7, 9] HeV has the capability of causing fatal diseases in several animal species including humans.[1] The primary host of the Hendra virus was identified as the flying fox species from the genus Pteropus[1,2,3] that resides and migrates through Northeastern Australia[8] or more specifically, the East coast of Australia to Melbourne and west across Northern Australia to Darwin[7].
Wang, Chih-Hung, et al. "Simian Virus 40 T Antigen Induces p53-Independent Apoptosis but does Not Suppress erbB2/neu Gene Expression in Immortalized Human Epithelial Cells." Cancer letters 137.1 (1999): 107-15.
Nowadays, technology makes easier for scientists to discover new viruses and find cures for them. One these viruses, “bird flu” known as H5N1 recreated by the Dutch scientist Ron Fouchier. Even though he had no ill intention to create such strain, it has a potential to wipe most humanity off from the face of the world as well as saving the humanity from much worse viruses. The discovery of this virus caused a big controversy in the scientific world. Some scientists think that publication of this virus indeed benefits society while some say that not publishing this might prevent society from horrible incidents.
Fanconi Anemia (FA) is a hereditary recessive disorder that is characterized by defective DNA cytogenetic instability, hypersensitivity to DNA crosslinking agents, increased chromosomal breakage, and cytogenetic instability. FA is caused by mutations in a complex set of proteins, including a FA core complex which contains eight out of sixteen known FA genes and their associated proteins. The FA proteins work together in a genome maintenance pathway called the FA/BRCA pathway, which plays an important role during the S phase of the cell cycle. The list FA complementation group (FANC) are: FANC-A, -B, -C, -D1/BRCA2, -D2, -E, -F, -G, -L, -I, -J/BRIP1,-M, -N/PALB2, -P/SLX4, -O/RAD51C and XPF. While the members of the FA complementation group do not share sequence similarity, they are related by their assembly into a common nuclear protein complex. Beside these sixteen FA proteins, there are several other proteins associating with the FA core complex, known as the FA Associated Proteins (FAAPs): -100, -24, -20, -16/MHF1, and -10/MHF2. FA plays an important role in the genomic stability through DNA repair of interstrand crosslinks (ICLs). When mutations occur in these genes, however, abnormal cell division, which eventually causes cancer and congenital defects occurs in most patients (Nalepa, et al., 2013; Tomida, et al., 2013).
Norovirus, also known as the "Norwalk Virus", is a highly contagious food-related illness that got its name when it was first discovered in Norwalk, Ohio in 19721. It is said to be frequently found in cruise ships, daycares and nursing homes that hold large amounts of people in close proximity2.Causes of Norovirus include a cook or bartender serving food or drink without washing their hands properly, having direct or close contact with someone is infected (touching them and then touching the mouth or nose) and drinking an infected person's water or using their utensils1. The very first cause listed is something an individual may not be able to control, which is why norovirus is so common and contagious.
...l electrophoresis (SCGE) also known as comet assay has become one of the standard methods for assessing DNA damage, with applications ranging from testing genotoxicity, human bio-monitoring and molecular epidemiology to its use in fundamental research in DNA damage and repair (Collins, 2004). The comet assay is a simple method for detecting DNA strand breaks within cells in eukaryotes. The procedure of comet assay includes Embedding the cells in agarose in a microscope slide, followed by lysing of cells with detergent and high salts to form nucleotides containing supercoiled loops of DNA linked to the nuclear matrix, and then undergoing Electrophoresis at high pH, which results in formation of structures resembling as comets, observed by fluorescence microscopy. The intensity of the comet tail relative to the head reflects the number of DNA breaks (Collins, 2004).
For approximately three-thousand years, smallpox has ravaged and plagued the four corners of the globe. In fact, in the 17 th and 18 th centuries, it was claimed to be the most infectious disease in the West, with an astounding 90% mortality rate in America. It wasn't until 1796, with English surgeon Edward Jenner's smallpox vaccination, that the world saw relief from this devastating virus. However, even with this inoculation in use, the world continued to witness death from both the virus and the vaccine. In the year 1966, it was estimated that 10-15 million infected citizens world wide had passed away from smallpox that year alone ( “History” 12). As a result of these devastating numbers, in the following year, 1967, the World Health Organization (W.H.O.) created a program to eradicate the smallpox virus. Ten years later, in 1977, the estimated 10-15 million cases had dwindled down to one; a man in Somalia. Three years later, W.H.O. officially announced that smallpox had been eradicated, leaving the only remaining virus cultures stored and guarded in laboratories in Russia and the United States. Inoculations ceased, smallpox epidemics were non-existent, and the virus was no longer a concern. In order to ensure complete eradication of this deadly virus, the W.H.O. insisted that the remaining smallpox cultures be destroyed by 1999 ( “Smallpox Eradication” 2). However, despite the W.H.O.'s recommendation, the remaining cultures continue to be contained and protected to this day, five years after the suggested date of elimination.
“This knowledge will help us design drugs that mimic the viral effects on these proteins to either activate a host’s immune response or shut it down,” said Dr. Michael Gale, associate ...
One class which belongs to the molecular chaperones is known as the chaperonins which form large, multisubunit cagelike structures in eubacteria, mitochondria, chloroplasts, and in the eukaryotic cytosol. The bacterial chaperonin is known as GroEL (L for large) and it performs its function along with its co-chaperonin GroES (S for small). Both of these are expressed from the GroE operon in Escherichia coli. In eukaryotes, the heat shock proteins Hsp60 and Hsp10 are homologous to GroEL and GroES, respectively, as they are almost identical in structure and function.
The birth of genetic engineering and recombinant DNA began in Stanford University, in the year 1970 (Hein). Biochemistry and medicine researchers were pursuing separate research pathways, yet these pathways converged to form what is now known as biotechnology (Hein). The biochemistry department was, at the time, focusing on an animal virus, and found a method of slicing DNA so cleanly that it would reform and go on to infect other cells. (Hein) The medical department focused on bacteria and developed a microscopic molecular messenger, that could not only carry a foreign “blueprint”, or message, but could also get the bacteria to read and copy the information. (Hein) One concept is needed to understand what happened at Stanford: how a bacterial “factory” turns “on” or “off”. (Hein) When a cell is dividing or producing a protein, it uses promoters (“on switches”) to start the process and terminators (“off switches”) to stop the process. (Hein) To form proteins, promoters and terminators are used to tell where the protein begins and where it ends. (Hein) In 1972 Herbert Boyer, a biochemist, provided Stanford with a bacterial enzyme called Eco R1. (Hein) This enzyme is used by bacteria to defend themselves against bacteriophages, or bacterial viruses. (Hein) The biochemistry department used this enzyme as a “molecular scalpel”, to cut a monkey virus called SV40. (Hein) What the Stanford researchers observed was that, when they did this, the virus reformed at the cleaved site in a circular manner. It later went on to infect other cells as if nothing had happened. (Hein) This proved that EcoR1 could cut the bonding sites on two different DNA strands, which could be combined using the “sticky ends” at the sites. (Hein). The contribution towards genetic engineering from the biochemistry department was the observations of EcoR1’s cleavage of
Marburg virus belongs to the genus Marburgvirus in the family Filoviridae, and causes a grave hemorrhagic fever, known as Marburg hemorrhagic fever (MHF), in twain humans and nonhuman primates. Basic Safety measures for medical personnel and others who are taking care of presumed individuals who may be contaminated with Marburg disease. Marburg Virus, Akin to the more widely known Ebola hemorrhagic fever, MHF is portrayed by systemic viral replication, lowering the body’s normal immune response to invasion by foreign substances and abnormal inflammatory responses. Ebola and Marburg Virus are very similar in many ways Marburg virus was introduced first in the 1960’s. These pathological features of the disease subsidize to a numerous of systemic dysfunctions including
These enzymes are widely used in numerous biotechnological processes and have applications in cosmetic, detergent, food, pharmaceutical industries and leather (Rajeshkumar, Mahendran et al. 2013).
With this information, of which is all theoretically possible, we now have a zombie that passes the principles of a zombie.The medical science involved has now proven that zombies are possible.Have fun and do not create this unless you want to be responsible for the mass genocide of the human race.