Tag: dna replication

DNA polymerase I serve clean-up functions during replication, recombination, and repair. It has 3' --> 5' proofreading exonuclease activity and 5' --> 3' exonuclease activity. The 5' --> 3' exonuclease activity of DNA polymerase I can replace an error containing the segment of DNA/RNA, paired to the template strand which is then simultaneously replaced by polymerase activity of the same enzyme. The process is called as nick translation. DNA polymerase III with its ten types of subunits has 5' --> 3' polymerization and 3' --> 5' proofreading activities in its $\alpha$ and $\epsilon$ subunits, respectively. 

During DNA replication, two hexamers of helicases are loaded onto each DNA strand. Helicases move along the double stranded DNA and separate the strands by breaking hydrogen bonds between base pairs. ATP hydrolysis provides the required energy for breaking of hydrogen bonds; options B and C are correct. Thereby, DNA is unwound bidirectionally and two potential replication forks are created. The point at which two DNA strands are separated to facilitate the replication is called replication fork; option A is correct. Correct answer is D.

Eukaryotes have three RNA polymerases which are structurally distinct complexes, though share certain subunits in common, and have a specific function and specific promoter sequence. RNA polymerase I synthesize preribosomal RNA (pre-rRNA), which contains the precursor for the 18S, 5.8S, and 28S rRNAs. RNA polymerase II is synthesized mRNAs and some specialized RNAs. RNA polymerase III makes tRNAs, the 5S rRNA, and some other small specialized RNAs. Option B is correct.

Two classes of enzymes are most important in generating recombinant DNA molecule. First, restriction endonucleases (also called restriction enzymes) recognize and cleave DNA at specific DNA sequences (recognition sequences or restriction sites) to generate a set of smaller fragments. Second, the DNA fragment to be cloned can be joined to a suitable cloning vector by using DNA ligases to link the DNA molecules together. DNA ligase catalyzes the formation of new phosphodiester bonds in a reaction that uses ATP or a similar cofactor.

DNA polymerase is an enzyme used in the replication of DNA. It ads the DNA primers to the parent chain. This enzyme acts in the 5'-3 direction and thus forms lading stand on the chain that has 3'5' direction.

RNA polymerase serves as primase during DNA replication and synthesizes RNA/DNA primer which is a stranded segment (complementary to the template) and has a free 3’ hydroxyl group to facilitate the addition of deoxyribonucleotides by DNA polymerase enzyme. DNA polymerase is polymerizing enzyme that has 5’ to 3’ polymerization and 3’ to 5’ exonuclease proofreading activities; it adds deoxyribonucleotides to free 3’ hydroxyl group of primer to synthesize new DNA strands. DNA ligase enzyme joins two Okazaki fragments together via phosphodiester bond between 3' hydroxyl at the end of one fragment and a 5' phosphate at the end of another fragment. Thus, option D is the correct answer. 

In molecular biology, DNA ligase is a specific type of enzyme, a ligase, that facilitates the joining of DNA strands together by catalyzing the formation of a phosphodiester bond.