6. You are studying a cargo protein which you believe is involved in nuclear tra
ID: 261589 • Letter: 6
Question
6. You are studying a cargo protein which you believe is involved in nuclear transport. You have created various truncation mutations in the cargo protein in which certain portions of are removed. You determine whether the cargo protein localizes to the nucleus and/or cytoplas conditions and also in the presence of leptomycin B, an inhibitor of the nuclear export machinery. Your data are shown in the following figure. (A plus means the cargo protein is located in that part of the cell, a minus that it isn't.) 51-750 m under normal 1-490 A. (4pts) Why does the removal of parts of this cargo protein (and others) cause the cargo protein to change its location in the cell? (4pts) How can you use this information to determine the location of a nuclear localization sequence (NLS) in this cargo protein? C. (2pts) What range of amino acids do you think contains the NLS? ES? _ D. (2pts) What range of amino acids do you think contains the N rus to determine how it infects host cells In the lab, you are investigating a newly identified vi Luckily, you obtained a GFP-expressing fluorescent endocyt ou c an observe infection via microscopy. You decide to use this tool to test whether clathrin-dependent osis is the required entry mechanism for this virus. virus from a colleague, so yExplanation / Answer
A) It is important to have NLS sequence on Cargo protein because without this the cargo protein cannot enter inside the nucleus. However, after entering in the nucleus, it must be release the NLS sequence because it has to free for other cargo proteins. Moreover, after performing its role, cargo protein again takes other sequences that help it to move outside. Therefore, it is a continuous process for differentiating and inserting the cargo proteins.
b) We can see that in normal condition, from 1- 750 and 151-750 amino acids the cargo protein is present at both place but in 301-750 amino acids, it is not present in cytoplasm. This is because this protein is now present in the nucleus and the NLS sequence is present from the 1-301 sequence. Now from the other result, we can see that 1-150 and 150-300 contains protein in cytoplasm while 151- 450 contains the cargo protein in both regions. It means the cargo protein is 300 amino acid long. Now, from 301 to 450 the protein is present in only in nucleus, here only NLS present.
C) the NLS present at 301- 450, 151-450, 1-750, 151- 750 1nd 301- 750.
D)Now the NES is present at the 1-150, 151-300, and 451-750 because the protein is present only in the cytoplasm. It also proved by the chart in presence of leptomycin.
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