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CryoEM structure of type IV-A CasDinG bound NTS-nicked Csf-crRNA-dsDNA quaternary complex in a second state

Electron Microscopy
3.7Å resolution
PDB entry 7xg4 coloured by chain, front view.
PDB entry 7xg4 coloured by chain, side view.
PDB entry 7xg4 coloured by chain, top view.
Source organism: Pseudomonas aeruginosa
Assembly composition:
hetero dodecamer (preferred)
Entry contents: 5 distinct polypeptide molecules
2 distinct DNA molecules
1 distinct RNA molecule

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Assemblies

Assembly 1 (preferred)
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Multimeric state: hetero dodecamer
Accessible surface area: 113624.19 Å2
Buried surface area: 65451.81 Å2
Dissociation area: 3,050.37 Å2
Dissociation energy (ΔGdiss): 26.28 kcal/mol
Dissociation entropy (TΔSdiss): 14.48 kcal/mol
Symmetry number: 1
PDBe Complex ID: PDB-CPX-222858

Macromolecules

Csf1
Chain: A
Length: 253 amino acids
Theoretical weight: 28.68 KDa
Source organism: Pseudomonas aeruginosa
Expression system: Escherichia coli
Csf1 in PDB entry 7xg4, assembly 1, front view.
Csf1 in PDB entry 7xg4, assembly 1, side view.
Csf1 in PDB entry 7xg4, assembly 1, top view.
125320406080100120140160180200220240
 
100200HHHHHHHHHHMRYPSDIVDQVLKAGPDKGLLTWEGVDAACSHCSRPIQNGDLYSPSSVGAFFSDTRDLASTSRSICWRCVVLRKKPMLYGLSAAVVTQDGIYSISKDVNKAWLFTTPPPAPFLVVHSSSTMQHLSWRTPVTLDNRRIHVRYGPNLFIVRPEVVRKALSIADRVNEGQKKWVTPVYFDRKAAAMGHGLITRAGAEMLTQEEQEFFQSVTPGERWALSYLMHSKRPEPEVGECITEKVMTSLNKE
Chains
Chain A (auth A)
Secondary structure
Ligand binding sites
Sequence conservation
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Csf3
Chain: B
Length: 220 amino acids
Theoretical weight: 24.42 KDa
Source organism: Pseudomonas aeruginosa
Expression system: Escherichia coli
Csf3 in PDB entry 7xg4, assembly 1, front view.
Csf3 in PDB entry 7xg4, assembly 1, side view.
Csf3 in PDB entry 7xg4, assembly 1, top view.
122020406080100120140160180200220
 
100200MVNLKVTIDLSNPMMEPGDLLHLDALLGALRVSRARAEHGDAINPRDYHYDLPLERYQAPSGDWVFKASAFKLKRQLPNQMWMQTGRLSIVEAARHRQSGYLQLRAGKPNPAGGPFKTSIYHRPIVQAELTAFCVGDQQGIEALLSECRQIGGKRGVGFGQVAGFKVEPVAETDCPWSWRALPADADPRLVTSEHARCIAAIRGPYWDRTLHVEALAPTP
Chains
Chain B (auth B)
Secondary structure
Sequence conservation
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Csf2
Chains: C, D, E, F, G
Length: 348 amino acids
Theoretical weight: 37.32 KDa
Source organism: Pseudomonas aeruginosa
Expression system: Escherichia coli
Csf2 in PDB entry 7xg4, assembly 1, front view.
Csf2 in PDB entry 7xg4, assembly 1, side view.
Csf2 in PDB entry 7xg4, assembly 1, top view.
134850100150200250300
 
100200300MQIEVTVRNITPIFSAAPGSNYITIDGTINPPPGVSRFPLVRTRMMYVAADVGDGVIKSVPLQIVPGNTMRSLLRRTMLKHVIEPALVEKGNKLSIGAYATAYSGNATGNPDGVPSSFDEIATMRAHPFIGLFGGGPRMLEGRLMVDSLYPIHTNAERILGAGYENEMMSGPITQVVWARRMDPILNLGSSEDVEVINGGAVAANGWIQDLLANSKAAASKKKKAAADEDESDGAAEENGRGLKAFNAHEVVIPGLKWVWRISLDRPTDAQVGLVLLALNKMTNERIAGGHSKDYGRFVIDGVSLNGEQVWSQSGITGGEQYFDAVAEAIDGLSSKEFEQFAQSAKEA
Chains
Chain C (auth C)
Chain D (auth D)
Chain E (auth E)
Chain F (auth F)
Chain G (auth G)
Secondary structure
Sequence conservation
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Csf5
Chain: H
Length: 268 amino acids
Theoretical weight: 29.15 KDa
Source organism: Pseudomonas aeruginosa
Expression system: Escherichia coli
Csf5 in PDB entry 7xg4, assembly 1, front view.
Csf5 in PDB entry 7xg4, assembly 1, side view.
Csf5 in PDB entry 7xg4, assembly 1, top view.
126820406080100120140160180200220240260
 
100200MFVKQVFFDLGARIHADEARALVAKLLDDTQPGLVSALMNYMPASKTSKTEFPLVQFSNFNQGFALLGFGEVGAQILSDATPIIHDAMAKLFASRGQGVVVQVSSRDVPLSCEKRPYGLQYTVAKMVVQKKHEHRERLANPETGKVFLEGLFLRSLERQAAAVGMVLPRDLVVSFKGAERVSSVKLRPDSTLAHGSLRHAVFEVNARLGGLWSVGFLLSKGFGHINTDLQLGQGGTRHALSKWSHPQFEKGGGSGGGSGGWSHPQFEK
Chains
Chain H (auth H)
Secondary structure
Sequence conservation
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Csf4
Chain: L
Length: 626 amino acids
Theoretical weight: 67.94 KDa
Source organism: Pseudomonas aeruginosa
Expression system: Escherichia coli
Csf4 in PDB entry 7xg4, assembly 1, front view.
Csf4 in PDB entry 7xg4, assembly 1, side view.
Csf4 in PDB entry 7xg4, assembly 1, top view.
162650100150200250300350400450500550600
 
200400600MELPADALPGEGAVREVLRPLLKQAAEKTAAGKIVFAEAATGTGKGRMIASLAAAAAIKGDTVVVSAPLAVTWQLVNDMKDIPEVRRVGLTLSLGRPNFISPQRTLEWAIDNERADLAAWIEGGGKPLSLRSMETSKVISHELCWLLEDALLLAEDLPADSLLLTSEDPADCPAQQLYVAMRSNYTEAGIILCSHFMLAAHTRMMQMRGLGNDEELDDEAPTGLSLPHFIDTLIVDEAHLLEQAFASVYTHTLRLRPLMRTIEGLGSRGRKPALDALKELFTQMQVASARSTNTSLNVPLSDVPELIPALKDTVKTLGALPTKGMSRDARSVIRIATRAANDALSGHSRLRIEVTPVHSYPMLLSGRSNLQRALLGLWNATGGATLVSATLFTTGDNGSLTRWKLEVPTERAAFLPPVHPAWTTAPVLLHKEFCAHEPDDSPEWATECAQTIQGVASTAQGGTLVLCTSYQNTELLAGRLGAALGDRLIVQSKTSSAATCLAQFKAKHKAGIRPVWLGLGAAWTGIDLSDHSLPDNPELDRLLSDLVITRIPVGQNRSLTHERRTAIGGFRIISQEAAWHFRQGLGRLVRRPGVTHKNLWVLDARIYGGAAWVAPFRQILDRYKKA
Chains
Chain L (auth L)
Domains
Secondary structure
Sequence conservation
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DNA chain J
Name: NTS
Representative chains: J
Length: 38 nucleotides
Theoretical weight: 11.52 KDa
NTS in PDB entry 7xg4, assembly 1, front view.
NTS in PDB entry 7xg4, assembly 1, side view.
NTS in PDB entry 7xg4, assembly 1, top view.
1385101520253035
 
102030AACACCCTTTCATTATTATTATTATTATTTTTTTTTTT
Chains
Chain J (auth J)
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DNA chain K
Name: TS
Representative chains: K
Length: 54 nucleotides
Theoretical weight: 16.55 KDa
TS in PDB entry 7xg4, assembly 1, front view.
TS in PDB entry 7xg4, assembly 1, side view.
TS in PDB entry 7xg4, assembly 1, top view.
1545101520253035404550
 
2040CTGCCGCACTTGCTCATCAAGCCTTCCTTCAGGTGTTGCTCCAGAAAGGGTGTT
Chains
Chain K (auth K)
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RNA chain I
Name: crRNA
Representative chains: I
Length: 61 nucleotides
Theoretical weight: 19.9 KDa
crRNA in PDB entry 7xg4, assembly 1, front view.
crRNA in PDB entry 7xg4, assembly 1, side view.
crRNA in PDB entry 7xg4, assembly 1, top view.
16151015202530354045505560
 
204060GUGAACGGUGGAGCAACACCUGAAGGAAGGCUUGAUGAGCGUGUUCCCCGCAUACGCGGG*
Chains
Chain I (auth I)
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