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[1mcataway]

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[unique135]

Various deletion mutations in Protein B are created and examination of its localization under normal conditions and also in the presence of "leptomycin B", an inhibitor of the nuclear export machinery.

This seems wrong.

 

[1mcataway]

I would have guessed both NLS and NES are in 240-357. This is obvious in NLS since it shows that in normal conditions this is the smallest deleted mutation that is responsible for its localization since it shows that this protein is present in the nucleus due to this part of the protein.
However, in NES I was not sure. I was using the same way I found it for NLS but I found it to be 1-114 and 115-239. But, these shouldn't be true because leptomycin B is supposedly inhibiting NES, thus this protein should not be located in the cytosol if leptomycin B is added and this protein portion has NES. 240-357 was the only smallest portion of the protein that has inhibited NES due to leptomycin because protein B was not present in the cytoplasm.
I don't know if I am going in the right direction.

 

[unique135]

I still can't comprehend the idea of mutation here.

There are four types of protein pieces:
1. that has both NLS and NES
2. that has only NLS
3. that has only NES
4. that has none

1-576, 115-576, 240-576, 115-357 and 240-357. In all these pieces, there is something common that drives these pieces in the nucleus.

1-576, 115-576, 115-357. In here, there is something that kicks out these pieces out of the nucleus.

Rest are those that either not driven in or kicked out.

 

[1mcataway]

I still can't comprehend the idea of mutation here.

There are four types of protein pieces:
1. that has both NLS and NES
2. that has only NLS
3. that has only NES
4. that has none

1-576, 115-576, 240-576, 115-357 and 240-357. In all these pieces, there is something common that drives these pieces in the nucleus.

1-576, 115-576, 115-357. In here, there is something that kicks out these pieces out of the nucleus.

Rest are those that either not driven in or kicked out.

I think it is asking for the minimal domain that the signals are in.
So I think
NES- 240-357
NLS- 115-357

 

[unique135]

http://i50.tinypic.com/2ymdmo7.jpg

Pieces with NLS are blue.
Pieces with NES are red.
Pieces that never entered (bcoz of missing NLS) are orange.

This should give you the answers.

 

[1mcataway]

http://i50.tinypic.com/2ymdmo7.jpg

Pieces with NLS are blue.
Pieces with NES are red.
Pieces that never entered (bcoz of missing NLS) are orange.

This should give you the answers.

Thanks for your help. I am confused on something though.
You pointed the region of the NES to be 115-239, however, in the presence of leptomycin, the NES is suppose to be inhibited. However, in the last case it isn't. What is the reason for this? I think the NES is suppose to be somewhere in the region between 115-357, in this case it is inhibited.

 

[unique135]

Thanks for your help. I am confused on something though.
You pointed the region of the NES to be 115-239, however, in the presence of leptomycin, the NES is suppose to be inhibited. However, in the last case it isn't. What is the reason for this? I think the NES is suppose to be somewhere in the region between 115-357, in this case it is inhibited.

Cystoplasmic proteins don't undergo post-translational modification. So, the journey of these proteins begins from cystoplasm. If protein has NLS & NES only then they can enter in the nucleus and come back out. If they had only NLS, they are unlikely to come out of the nucleus. If they had NES only, they will never go in the nucleus. In 115-239, this piece was lacking NLS (ticket to enter nucleus) so it will never go in and remain in cytosoplasm.

I think the NES is suppose to be somewhere in the region between 115-357, in this case it is inhibited.

Yes, NES is in 115-357. From the question setup, it is 115-239 (115-357 is not in options). Apart, it could be that NES is half in 115-239 and half in 240-357. There is no way to find out from this data. NES is ~10aa so what are the chances that NES overlaps with NLS.