7. DNA and Chromosome Structure

In this chapter, we focused on how DNA is organized at the chromosomal level. Along the way, we found many opportunities to consider the methods and reasoning by which much of this information was acquired. From the explanations given in the chapter, what answers would you propose to the following fundamental questions:How do we know that viral and bacterial chromosomes most often consist of circular DNA molecules devoid of protein?

Write a short essay that contrasts the major differences between the organization of DNA in viruses and bacteria versus eukaryotes.

Contrast the size of the single chromosome in bacteriophage and T2 with that of E. coli. How does this relate to the relative size and complexity of phages and bacteria?

Assume that a viral DNA molecule is a 50-µm-long circular strand with a uniform 20-Å diameter. If this molecule is contained in a viral head that is a 0.08-µm-diameter sphere, will the DNA molecule fit into the viral head, assuming complete flexibility of the molecule? Justify your answer mathematically.

How many base pairs are in a molecule of phage T2 DNA 52-µm long?

Following is a diagram of the general structure of the bacteriophage chromosome. Speculate on the mechanism by which it forms a closed ring upon infection of the host cell.

To gauge the fidelity of DNA synthesis, Arthur Kornberg and colleagues devised a technique called nearest-neighbor analysis, which determines the frequency with which any two bases occur adjacent to each other along the polynucleotide chain (J. Biol. Chem. 236: 864–875). This test relies on the enzyme spleen phosphodiesterase (see the previous problem). As we saw in Figure 11-8, DNA is synthesized by polymerization of 5'-nucleotides—that is, each nucleotide is added with the phosphate on the of deoxyribose. However, as shown in the accompanying figure, the phosphodiesterase enzyme cleaves DNA between the phosphate and the C-5' atom, thereby producing 3'-nucleotides. In this test, the phosphates on only one of the four nucleotide precursors of DNA (cytidylic acid, for example) are made radioactive with ³²P and DNA is synthesized. Then the DNA is subjected to enzymatic cleavage, in which the radioactive phosphate is transferred to the base that is the 'nearest neighbor' on the 5' side of all cytidylic acid nucleotides.Following four separate experiments, in each of which a different one of the four nucleotide types is radioactive, the frequency of all 16 possible nearest neighbors can be calculated. When Kornberg applied the nearest-neighbor frequency test to the DNA template and resultant product from a variety of experiments, he found general agreement between the nearest-neighbor frequencies of the two. Analysis of nearest-neighbor data led Kornberg to conclude that the two strands of the double helix are in opposite polarity to one another. Demonstrate this approach by determining the outcome of such an analysis if the strands of DNA shown here are (a) antiparallel versus (b) parallel: