Question

You have identified a novel activator protein. You assume that like other activators, your protein has activation and DNA binding domains. You carry out deletion analysis on the part of the ORF that codes for the C-terminal end of your activator and assay both DNA binding and transcription. You obtain the results indicated in the figure below. Answer the following 3 parts (i,ii, & iii) based on this information.

Txn +++ DNA binding + 1 Activator ORF 600 1 550 +++ + 1 500 +++ + 1 450 +++ + 1 400 + 1 350 + 1 + 300 1 250 + 1 200 +

i. Which most accurately describes which part of the protein contains the DNA binding domain?

A. It contained in the first 200 amino acids

B. It is contained in amino acids 250-350

C. It is contained in amino acids 300-400

D. It is contained in amino acids 350-400

E. It is contained in amino acids 300-450

F. It is contained in amino acids 350-450

G. It is contained in amino acids 450-600

ii. Which most accurately describes which part of the protein contains an activation domain?

A. It contained in the first 200 amino acids

B. It is contained in amino acids 250-350

C. It is contained in amino acids 300-400

D. It is contained in amino acids 350-400

E. It is contained in amino acids 300-450

F. It is contained in amino acids 350-450

G. It is contained in amino acids 450-600

You continue your analysis of this activator protein and construct a series of deletion mutants in which segments in the middle part of the protein are missing. You obtain the data shown in the figure below.

iii. What can you conclude from this data?

(ans) That the protein contains two activation domains. One activation domain is located between amino acids 350 and 450, and that the other activation domain is located between amino acids 450 and 550.

Answer 1

I) the DNA binding domain is represented by amino acids which are before aminos code 200 because after deletion of all these amino acids after 200 to 600 has not affect the DNA binding so, this domain represent DNA binding domain.

ii)the right answer will be the aminos code from 350 to 450 because transcription of target gene become less after deletion of this amino acids regions.

Other deletion has not affect the Transcription.

Iii)you have determined the thing is logical but your value is different like these have two domain but the responsible Amino acids will be

350-400 and 450- 500 because you can see,

In deletion 5,deletion from 200 to 350 has no Transcription effects but in deletion 4, deletion from 200 to 400 has huge effects, so the crucial amino acids is 350 to 400.

And , if you see deletion 3 and 4, then you will see that amino acids 450-500 is responsible similarly.

Thank you