Kutay_guide_answers - Study Guide Kutay et al. (1997)...

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Study Guide Kutay et al. (1997) Export of importin α from the nucleus is mediated by a specific nuclear transport factor. Cell 90: 1061-1071. These questions are intended to help you read the paper in an inquisitive fashion and to help you focus on some of the issues you should understand. Please cover each of these questions during your reading and studying. Please submit the answers to 5 of these questions for your homework assignment. Questions 1) We discussed nuclear import of proteins with a classic NLS in lecture 4. A fundamental problem of the nuclear import process is that it is an asymmetric cycle, after the import receptors bring cargo to the nucleus, the import receptors must be re-exported. What are the authors labeling an “exportin”, and what is the specific role of the exportin called CAS that they have identified in this paper? Exportin is a nuclear export receptor and CAS is the export receptor for importin α 2) In Figure 1, the authors are monitoring import of a specific nuclear cargo (nuceloplasmin) and then re- export of importin α . a. Is the nuclear envelope intact in this assay? b. Is the cytoplasmic membrane intact in this assay? c. BRIEFLY outline how are they monitoring the position (nuclear versus cytoplasmic) of importin α ? d. How do they know that a round of import of nuclear cargo took place? a. yes b. no c. importin α is labeled with a fluorescent moiety – fluorescein. They monitor its position using a confocal microscope (laser equipped microscope capable of exciting the fluorescein in a single plane at a time, making it easy to determine whether fluorescence is nuclear or cytoplasmic)
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Kutay_guide_answers - Study Guide Kutay et al. (1997)...

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