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FnCas9 major
domains, colored by domain:
Cyan
for RuvC, cleaves the non-target DNA strand
Green
for BH (bridge helix), spans the channel
Light
gray for REC1
Black
for REC2
Dark
gray for REC3
Pink for
HNH, cleaves the target DNA strand
Gold for
Wed
Khaki
for PI
DNA
purple, sgRNA magenta
Highlights
residues along the main channel and DNA/RNA entry channels.
The RuvC-I
subdomain (cyan) is attached to REC, opposite the rest of RuvC
(purple), and must join RuvC by using the bridge helix (green) to
span the channel. RuvC-I is primarily a beta-sheet, and fits into
a RuvC pocket in line with two other RuvC beta-strands (gold).
DNA (purple) and
the sgRNA (magenta) in and around FnCas9. DNA enters from between
PI (khaki) and WED (gold), where the target strand splits from the
non-target strand. The target strand continues upward and joins
one end of the sgRNA, which enters from WED, and together they
form a DNA:RNA duplex in the main channel. The back end of the
sgRNA forms a repeat:antirepeat structure sticking out of WED, and
then wraps around the back of the REC domains.
The TGG PAM (T in
brown and G's in gold), and three residues in FnCas9's PI domain
responsible for recognizing the PAM: arginines 1556 and 1585
(green), and serine 1473 (blue). Hydrogen-bonding shown in red.
H. Hirano, J.S. Gootenberg, T. Horii, O.O.
Abudayyeh, M. Kimura, P.D. Hsu, T. Nakane, R. Ishitani, I.
Hatada, F. Zhang, H. Nishimasu, O. Nureki.
“Structure
and Engineering of Francisella novicida Cas9.”
Cell 164.5 (2016): 950-961 (PubMed)
5b2o (PDB)
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