How would you prove experimentally that the RNA polymerase goes in only one direction (5’ to 3’) and not the other way?
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I would like to clarify (even though the question mentions it) that I have asked for an "Experimental" Procedure. Theory we all know. The question is how we would Experimentally prove this fact that RNA Polymerase goes only in the 5' to 3' end, and not the other way round.
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These fragments are processed by the replication machinery to produce a continuous strand of DNA and hence a complete daughter DNA helix. DNA replication goes in the 5' to 3' direction because DNA polymerase acts on the 3'-OH of the existing strand for adding free nucleotides.
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ANSWER
DNA-polymerase can only work from the 5'-end to the 3'-end.
Because of the structure of a nucleotide.
1) A nucleotide has two ends=>
i)free 5' phosphate end, ii)A free 3' OH end.
2) The strand in 5' to 3' direction indicates that an free 5' phosphate is present at one end and the other free 3' OH at the other end.
3) Nucleotides monomers are to be added to the 3’ OH end of the growing strand one by one by DNA Polymerase. Therefore new polynucleotide chain is always synthesised in the 5’-3’ direction.
4) DNA pol III involved in replication cannot add the first nucleotide to start the synthesise of daughter DNA strand.
5)DNA polymerase cannot initiate a synthesis but it requires a primer called primase, an RNA polymerase.
6)It synthesises a primer or a short sequence of around 10 RNA nucleotides complementary to parental DNA strand. It is also known as primer RNA. DNA pol III recognises the primer that provides a free 3'OH end and adds DNA nucleotide to construct a new DNA strand.
7)Later the RNA primer is replaced by an DNA nucleotides by DNA pol I.
DNA-polymerase can only work from the 5'-end to the 3'-end.
Because of the structure of a nucleotide.
1) A nucleotide has two ends=>
i)free 5' phosphate end, ii)A free 3' OH end.
2) The strand in 5' to 3' direction indicates that an free 5' phosphate is present at one end and the other free 3' OH at the other end.
3) Nucleotides monomers are to be added to the 3’ OH end of the growing strand one by one by DNA Polymerase. Therefore new polynucleotide chain is always synthesised in the 5’-3’ direction.
4) DNA pol III involved in replication cannot add the first nucleotide to start the synthesise of daughter DNA strand.
5)DNA polymerase cannot initiate a synthesis but it requires a primer called primase, an RNA polymerase.
6)It synthesises a primer or a short sequence of around 10 RNA nucleotides complementary to parental DNA strand. It is also known as primer RNA. DNA pol III recognises the primer that provides a free 3'OH end and adds DNA nucleotide to construct a new DNA strand.
7)Later the RNA primer is replaced by an DNA nucleotides by DNA pol I.
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