Bacteriophage T7 encodes its own DNA polymerase, gp5, which has both polymerase activity and exonuclease activity. Researchers made two sets of mutations in gp5, mutating various aspartate residues to alanine residues. The mutations are D5A/D65A (blue), D475A/D654A (orange) and D5A/D65A/D475A/D654A (green). They then used isotopically labeled dNTPs to quantify the activity of these mutants compared to wild type gp5 (red). The assay results (A and B) are shown below: A dGMP incorp. (pmol) 30 20 (b) D5A/D65A (c) WT DSA/DISA/DH75A/DISHA D475A/DESTA 500 1000 1500 2000 [gp5] (nM) dNMP released/min (pmol) 20 WT DV75A/DESHA DSA/DA/D47A/DUHA OSA/DISA 11 0 100 200 300 400 500 [gp5] (nM) 10 (a) Which data figure is measuring the polymerase activity? Which data figure is measuring the exonuclease activity? To measure the polymerase activity, which phosphate in the dNTPs needed to be isotopically labeled? Based on the data results, which mutations made the polymerase activity dysfunctional? Which mutations made the exonuclease activity dysfunctional?

Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
icon
Related questions
Question

please explain ALL parts of this one question clearly and in depth!

Bacteriophage T7 encodes its own DNA polymerase, gp5, which has both polymerase activity and
exonuclease activity. Researchers made two sets of mutations in gp5, mutating various aspartate residues
to alanine residues. The mutations are D5A/D65A (blue), D475A/D654A (orange) and
D5A/D65A/D475A/D654A (green). They then used isotopically labeled dNTPs to quantify the activity of
these mutants compared to wild type gp5 (red). The assay results (A and B) are shown below:
A
dGMP incorp. (pmol)
30
(b)
D5A/D65A
(c)
WT
DSA/DISA/DH75A/D6STA
D475A/D651A
500 1000 1500 2000
[gp5] (nm)
B
dNMP released/min (pmol)
50
40
30
20
WT
D475654
DSA/DA/D47A/DUSHA
OSA/DISA
4
0 100 200 300 400 500
[gp5) (nm)
10
(a)
Which data figure is measuring the polymerase activity? Which data figure is measuring
the exonuclease activity?
To measure the polymerase activity, which phosphate in the dNTPs needed to be
isotopically labeled?
Based on the data results, which mutations made the polymerase activity dysfunctional?
Which mutations made the exonuclease activity dysfunctional?
Transcribed Image Text:Bacteriophage T7 encodes its own DNA polymerase, gp5, which has both polymerase activity and exonuclease activity. Researchers made two sets of mutations in gp5, mutating various aspartate residues to alanine residues. The mutations are D5A/D65A (blue), D475A/D654A (orange) and D5A/D65A/D475A/D654A (green). They then used isotopically labeled dNTPs to quantify the activity of these mutants compared to wild type gp5 (red). The assay results (A and B) are shown below: A dGMP incorp. (pmol) 30 (b) D5A/D65A (c) WT DSA/DISA/DH75A/D6STA D475A/D651A 500 1000 1500 2000 [gp5] (nm) B dNMP released/min (pmol) 50 40 30 20 WT D475654 DSA/DA/D47A/DUSHA OSA/DISA 4 0 100 200 300 400 500 [gp5) (nm) 10 (a) Which data figure is measuring the polymerase activity? Which data figure is measuring the exonuclease activity? To measure the polymerase activity, which phosphate in the dNTPs needed to be isotopically labeled? Based on the data results, which mutations made the polymerase activity dysfunctional? Which mutations made the exonuclease activity dysfunctional?
Expert Solution
steps

Step by step

Solved in 2 steps

Blurred answer
Similar questions
  • SEE MORE QUESTIONS
Recommended textbooks for you
Biochemistry
Biochemistry
Biochemistry
ISBN:
9781319114671
Author:
Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:
W. H. Freeman
Lehninger Principles of Biochemistry
Lehninger Principles of Biochemistry
Biochemistry
ISBN:
9781464126116
Author:
David L. Nelson, Michael M. Cox
Publisher:
W. H. Freeman
Fundamentals of Biochemistry: Life at the Molecul…
Fundamentals of Biochemistry: Life at the Molecul…
Biochemistry
ISBN:
9781118918401
Author:
Donald Voet, Judith G. Voet, Charlotte W. Pratt
Publisher:
WILEY
Biochemistry
Biochemistry
Biochemistry
ISBN:
9781305961135
Author:
Mary K. Campbell, Shawn O. Farrell, Owen M. McDougal
Publisher:
Cengage Learning
Biochemistry
Biochemistry
Biochemistry
ISBN:
9781305577206
Author:
Reginald H. Garrett, Charles M. Grisham
Publisher:
Cengage Learning
Fundamentals of General, Organic, and Biological …
Fundamentals of General, Organic, and Biological …
Biochemistry
ISBN:
9780134015187
Author:
John E. McMurry, David S. Ballantine, Carl A. Hoeger, Virginia E. Peterson
Publisher:
PEARSON