Background You are working on a new antibiotic. You suspect that this antibiotic

Question

Background You are working on a new antibiotic. You suspect that this antibiotic targets the prokaryotic (bacterial) replication machinery. To test this, you set up a replication experiment. You add al the components needed for replication (proteins and ribonucleic acids), an original DNA with all the necessary replication sequences, and radioactively labeled deoxyribonucleotides (dNTPs) to two test tubes, and then add your drug to the experimental tube, but not the control tube Because the only dNTPs available for replication are radioactively labeled, all new DNA generated in your replication experiment will be radioactive (hot). You hypothesize that if your drug blocks replication, you will not see any hot plasmid DNA in your experimental tube PART 1 To distinguish your original DNA from products of new replication, you decide to add nucleotides labeled with radioactive phosphate (P32) to the mix. Refer to the picture below. Can you use nucleotides that are labeled with P32 at the y location (y-P32)? At the a location (a-P32 )? Both? Why or why not? Phosphates 0 H2c o base 3, Deoxyribose sugar

Explanation / Answer

We will only use alpha position because the beta- and gamma-phosphates are lost because the alpha-phosphate is required for chain elongation, it evolved that way and those are the reactions that the various polymerases catalyze, and it makes thermodynamic sense to release pyrophosphate to power entire process while leaving the more stable nucleotide monophosphate in the nucleic acid backbone.

in dna and rna polymerization, the 5' phosphate group is required for chain elongation. the three' oh group of 1 nucleotide hooks onto the 5' phosphate institution of the adjacent nucleotide. the sugar-phosphate backbone of nucleic acid chains includes the alpha-phosphate group.

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