13. Gene Regulation in Eukaryotes

Overview of Eukaryotic Gene Regulation

Open Question
The Drosophila even-skipped (eve) gene is expressed in seven stripes in the segmentation pattern of the embryo. A sequence segment of 8 kb 5' to the transcription start site (shown as in the accompanying figure) is required to drive expression of a reporter gene (lacZ) in the same pattern as the endogenous eve gene. Remarkably, expression of most of the seven stripes appears to be specified independently, with stripe 2 expression directed by regulatory sequences in the region 1.7 kb 5' to the transcription start site. To further examine stripe 2 regulatory sequences, you create a series of constructs, each containing different fragments of the 1.7-kb region of the 5' sequence. In the lower part of the figure, the bars at left represent the sequences of DNA included in your reporter gene constructs, and the + and − signs at right indicate whether the corresponding eve-lacZ reporter gene directs stripe 2 expression in Drosophila embryos transformed through P element mediation. How would you interpret the results—that is, where do the regulatory sequences responsible for stripe 2 expression reside?
Open Question
RNA helicases are a class of proteins that bind mRNAs and influence their secondary structures and interactions with other proteins. RNA helicases have been implicated in many steps of RNA regulation such as splicing, decay, and translation. Why might these enzymes be so ubiquitously required for RNA regulation?
Open Question
During an examination of the genomic sequences surrounding the human β-globin gene, you discover a region of DNA that bears sequence resemblance to the glucocorticoid response element (GRE) of the human metallothionein IIA (hMTIIA) gene. Describe experiments that you would design to test (1) whether this sequence was necessary for accurate β-globin gene expression and (2) whether this sequence acted in the same way as the hMTIIA gene's GRE.
Open Question
Marine stickleback fish have pelvic fins with long spines that provide protection from larger predatory fish. Some stickleback fish were trapped in lakes and have adapted to life in a different environment. Many lake populations of stickleback fish lack pelvic fins. Shapiro et al. (2004) (Nature 428:717.723) mapped the mutation associated with the loss of pelvic fins to the Pitx1 locus, a gene expressed in pelvic fins, the pituitary gland, and the jaw. However, the coding sequence of the Pitx1 gene is identical in marine and lake stickleback [Chan et al. (2010). Science 327:5963,302–305]. Moreover, when the Pitx1 coding region is deleted, the fish die with defects in the pituitary gland and the jaw, and they lack pelvic fins. Explain how a mutation near, but outside of, the coding region of Pitx1 may cause a loss of pelvic fins without pleiotropic effects on the pituitary gland and jaw.
Open Question
The localization and translational control of actin mRNA is important for the migration of fibroblasts and is regulated by the activity of the kinase Src (see Figure 18.10). Src is activated by phosphorylation when cell surface receptors bind to signaling molecules. How might this system lead to a cell migrating in a specific direction? How might the cell migrate away from repulsive signals?