13. Gene Regulation in Eukaryotes

From the data in Table 19.3, draw up a list of histone H3 modifications associated with gene activation. Then draw up a list of H3 modifications associated with repression.

Are there any overlaps on the lists?

Microbiologists describe the processes of transcription and translation as 'coupled' in bacteria. This term indicates that a bacterial mRNA can be undergoing transcription at the same moment it is also undergoing translation. Is coupling of transcription and translation possible in single-celled eukaryotes such as yeast? Why or why not?

Amino acids are classified as positively charged, negatively charged, or electrically neutral.

How does acetylation of lysine affect its interaction with DNA, and how is this related to the activation of gene expression?

Amino acids are classified as positively charged, negatively charged, or electrically neutral.

How does this property of lysine allow it to interact with DNA?

Amino acids are classified as positively charged, negatively charged, or electrically neutral.

Which category includes lysine?

A full-length eukaryotic gene is inserted into a bacterial chromosome. The gene contains a complete promoter sequence and a functional polyadenylation sequence, and it has wild-type nucleotides throughout the transcribed region. However, the gene fails to produce a functional protein. What changes would you recommend to permit expression of this eukaryotic gene in a bacterial cell?

A full-length eukaryotic gene is inserted into a bacterial chromosome. The gene contains a complete promoter sequence and a functional polyadenylation sequence, and it has wild-type nucleotides throughout the transcribed region. However, the gene fails to produce a functional protein. List at least three possible reasons why this eukaryotic gene is not expressed in bacteria.

Methylation of H3K9 by itself silences genes, but if H3K4 and H4K20 are also methylated, the combination of modifications stimulates transcription. What conclusions can you draw about this?

A particular type of anemia in humans, called β-thalassemia, results from a severe reduction or absence of the normal β-globin chain of hemoglobin. However, the γ-globin chain, normally only expressed during fetal development, can functionally substitute for β-globin. A variety of studies have explored the use of the nucleoside 5-azacytidine for the expression of γ-globin in adult patients with β-thalassemia.

How might 5-azacytidine lead to expression of γ-globin in adult patients?

A particular type of anemia in humans, called β-thalassemia, results from a severe reduction or absence of the normal β-globin chain of hemoglobin. However, the γ-globin chain, normally only expressed during fetal development, can functionally substitute for β-globin. A variety of studies have explored the use of the nucleoside 5-azacytidine for the expression of γ-globin in adult patients with β-thalassemia.

Explain why this drug may also have some adverse side effects.

DNA methylation is commonly associated with a reduction of transcription. The following data come from a study of the impact of the location and extent of DNA methylation on gene activity in eukaryotic cells. A bacterial gene, luciferase, was inserted into plasmids next to eukaryotic promoter fragments. CpG sequences, either within the promoter and coding sequence (transcription unit) or outside of the transcription unit, were methylated to various degrees, in vitro. The chimeric plasmids were then introduced into cultured cells, and luciferase activity was assayed. These data compare the degree of expression of luciferase with differences in the location of DNA methylation [Irvine et al. (2002). Mol. and Cell. Biol. 22:6689–6696]. What general conclusions can be drawn from these data?