I-SceI

Type

Wild-Type


Target Sequence

-11  -10  -9   -8   -7   -6   -5   -4   -3   -2   -1   1    2    3    4    5    6    7    8    9    10   11  
A C G C T A G G G A T A A C A G G G T A A T

FASTA Sequence



MKNIKKNQVMNLGPNSKLLKEYKSQLIELNIEQFEAGIGLILGDAYIRSRDEGKTYCMQFEWKNKAYMDHVCLLY
DQWVLSPPHKRERVNHLGNLVITWTAQTFKHQAFNKLANLFIVNNKKTIPNNLVENYLTPMSLAYWFMDDGGKWD
YNKNSTNKSIVLNTQSFTFEEVEYLVKGLRNKFQLNCYVKINKNKPIIYIDSMSYLIFYNLIKPYLIPQMMYKLP
NTISSETFLK

Crystal Structure

PDB File
PyMOL File

Position Weight Matrix

I-SceI
posACGT
-11 0.25 0.25 0.25 0.25
-10 0.25 0.25 0.25 0.25
-9 0.25 0.25 0.25 0.25
-8 0.25 0.25 0.25 0.25
-7 0.10 0.05 0.05 0.80
-6 0.60 0.20 0.15 0.05
-5 0.06 0.12 0.70 0.12
-4 0.00 0.07 0.85 0.08
-3 0.00 0.10 0.80 0.10
-2 0.95 0.00 0.05 0.00
-1 0.00 0.28 0.00 0.72
1 0.70 0.00 0.32 0.00
2 0.65 0.10 0.25 0.00
3 0.00 1.00 0.00 0.00
4 0.65 0.35 0.00 0.00
5 0.08 0.08 0.76 0.08
6 0.00 0.08 0.92 0.00
7 0.10 0.10 0.40 0.40
8 0.07 0.50 0.07 0.36
9 0.30 0.10 0.30 0.30
10 0.45 0.15 0.20 0.20
11 0.40 0.07 0.08 0.45

Specificity Changing Mutations

Amino Acid Mutations (?)                 Specificity Changes                                          Reference
K86R, G100T +4A to +4C, +4C to +4C 1
K86R, G100C +4A to +4C, +4C to +4C 1
K86R, G100S +4A to +4C, +4C to +4C 1
W149G, D150C, N152K -7T to -7C 2

1     Joshi et al. (2011) Evolution of I-SceI homing endonucleases with increased DNA recognition site specificity. J. Mol. Biol., 405, 185-200.
2     Doyon et al. (2006) Directed evolution and substrate specificity profile of homing endonuclease I-SceI. JACS, 128, 2477-2484.


Additional Information

Author: Barry Stoddard

Notes:


Date Added: 2011-10-18