The protein NK2 is a transcriptional regulator that functions both as a repressor and activator. To map the two domains more precisely a research group introduced deletion mutations within the coding sequences and studied the effect of a mutation on the NK2 regulatory function. A reporter construct (b-gal activity measurement) was used to monitor gene expression (row 15).  Expression in the absence of NK2 was set at 100% (row 15). Repression is indicated by a significant reduction in expression (see row 14 for example) and activation is indicated by higher that 100% activity (e. g., row 11).  The presence of the wild type NK2 led to near complete inhibition of expression (row 14). In the figure a deletion is shown as an open area and the residues deleted are indicated on the left (e.g., the row 13 mutant has a deletion of residues 1-97). - a)    Which region(s) of the protein is required for repression function? - b)    Is the C-terminal end required for repression? - c)    What is the minimum region needed for repression? - d)    Is the region labelled ‘HD’ sufficient for activation? - e)    What is the minimum region needed for activation? (A region refers to an amino acid segment, e.g., aa 15-275)

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Chapter12: Commercial Insurance
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The protein NK2 is a transcriptional regulator that functions both as a repressor and activator. To map the two domains more precisely a research group introduced deletion mutations within the coding sequences and studied the effect of a mutation on the NK2 regulatory function. A reporter construct (b-gal activity measurement) was used to monitor gene expression (row 15). 

Expression in the absence of NK2 was set at 100% (row 15). Repression is indicated by a significant reduction in expression (see row 14 for example) and activation is indicated by higher that 100% activity (e. g., row 11).  The presence of the wild type NK2 led to near complete inhibition of expression (row 14). In the figure a deletion is shown as an open area and the residues deleted are indicated on the left (e.g., the row 13 mutant has a deletion of residues 1-97).

- a)    Which region(s) of the protein is required for repression function?

- b)    Is the C-terminal end required for repression?

- c)    What is the minimum region needed for repression?

- d)    Is the region labelled ‘HD’ sufficient for activation?

- e)    What is the minimum region needed for activation?

(A region refers to an amino acid segment, e.g., aa 15-275)

 

1 RD-Hom
-234567890123
11
12
13
14
15
A(167-277)
A(194-277)
A(97-138)
A(97-167)
A(97-194)
N595A
A(619-723)
HD
HA
I
I
A(1-392)
A(1-277) I
A(1-194) I
A(1-97) I
Wt NK-2
pUC41-75hsp40 Bgal
0
HD
100
150
200
Normalized % of B-gal activity
50
250
Figure 1. Effect of deletion in NK-2 on its transcription regulatory functions. The
rectangles identify the segments present. Sequences deleted are shown as open areas.
Transcribed Image Text:1 RD-Hom -234567890123 11 12 13 14 15 A(167-277) A(194-277) A(97-138) A(97-167) A(97-194) N595A A(619-723) HD HA I I A(1-392) A(1-277) I A(1-194) I A(1-97) I Wt NK-2 pUC41-75hsp40 Bgal 0 HD 100 150 200 Normalized % of B-gal activity 50 250 Figure 1. Effect of deletion in NK-2 on its transcription regulatory functions. The rectangles identify the segments present. Sequences deleted are shown as open areas.
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