HEADER VIRAL PROTEIN 16-FEB-05 1YVX
TITLE HEPATITIS C VIRUS RNA POLYMERASE GENOTYPE 2A IN COMPLEX
TITLE 2 WITH NON- NUCLEOSIDE ANALOGUE INHIBITOR
COMPND MOL_ID: 1;
COMPND 2 MOLECULE: RNA DEPENDENT RNA POLYMERASE;
COMPND 3 CHAIN: A;
COMPND 4 ENGINEERED: YES;
COMPND 5 OTHER_DETAILS: GENOTYPE 2A
SOURCE MOL_ID: 1;
SOURCE 2 ORGANISM_SCIENTIFIC: HEPATITIS C VIRUS;
SOURCE 3 ORGANISM_TAXID: 11103;
SOURCE 4 EXPRESSION_SYSTEM: ESCHERICHIA COLI BL21(DE3);
SOURCE 5 EXPRESSION_SYSTEM_TAXID: 469008;
SOURCE 6 EXPRESSION_SYSTEM_STRAIN: BL21(DE3);
SOURCE 7 EXPRESSION_SYSTEM_VECTOR_TYPE: PLASMID;
SOURCE 8 EXPRESSION_SYSTEM_PLASMID: PET-21B
KEYWDS NS5B POLYMERASE GENOTYPE 2A, NON-NUCLEOSIDE INHIBITOR,
KEYWDS 2 VIRAL PROTEIN
EXPDTA X-RAY DIFFRACTION
AUTHOR B.K.BISWAL,M.M.CHERNEY,M.WANG,L.CHAN,C.G.YANNOPOULOS,
AUTHOR 2 D.BILIMORIA,O.NICOLAS,J.BEDARD,M.N.G.JAMES
REVDAT 3 24-FEB-09 1YVX 1 VERSN
REVDAT 2 10-MAY-05 1YVX 1 JRNL
REVDAT 1 22-MAR-05 1YVX 0
JRNL AUTH B.K.BISWAL,M.M.CHERNEY,M.WANG,L.CHAN,
JRNL AUTH 2 C.G.YANNOPOULOS,D.BILIMORIA,O.NICOLAS,J.BEDARD,
JRNL AUTH 3 M.N.G.JAMES
JRNL TITL CRYSTAL STRUCTURES OF THE RNA DEPENDENT RNA
JRNL TITL 2 POLYMERASE GENOTYPE 2A OF HEPATITIS C VIRUS REVEAL
JRNL TITL 3 TWO CONFORMATIONS AND SUGGEST MECHANISMS OF
JRNL TITL 4 INHIBITION BY NON-NUCLEOSIDE INHIBITORS.
JRNL REF J.BIOL.CHEM. V. 280 18202 2005
JRNL REFN ISSN 0021-9258
JRNL PMID 15746101
JRNL DOI 10.1074/JBC.M413410200
REMARK 1
REMARK 2
REMARK 2 RESOLUTION. 2.00 ANGSTROMS.
REMARK 3
REMARK 3 REFINEMENT.
REMARK 3 PROGRAM : CNS 1.1
REMARK 3 AUTHORS : BRUNGER,ADAMS,CLORE,DELANO,GROS,GROSSE-
REMARK 3 : KUNSTLEVE,JIANG,KUSZEWSKI,NILGES, PANNU,
REMARK 3 : READ,RICE,SIMONSON,WARREN
REMARK 3
REMARK 3 REFINEMENT TARGET : NULL
REMARK 3
REMARK 3 DATA USED IN REFINEMENT.
REMARK 3 RESOLUTION RANGE HIGH (ANGSTROMS) : 2.00
REMARK 3 RESOLUTION RANGE LOW (ANGSTROMS) : 38.69
REMARK 3 DATA CUTOFF (SIGMA(F)) : 0.000
REMARK 3 DATA CUTOFF HIGH (ABS(F)) : 2945543.830
REMARK 3 DATA CUTOFF LOW (ABS(F)) : 0.0000
REMARK 3 COMPLETENESS (WORKING+TEST) (%) : 99.3
REMARK 3 NUMBER OF REFLECTIONS : 55391
REMARK 3
REMARK 3 FIT TO DATA USED IN REFINEMENT.
REMARK 3 CROSS-VALIDATION METHOD : THROUGHOUT
REMARK 3 FREE R VALUE TEST SET SELECTION : RANDOM
REMARK 3 R VALUE (WORKING SET) : 0.209
REMARK 3 FREE R VALUE : 0.244
REMARK 3 FREE R VALUE TEST SET SIZE (%) : 5.100
REMARK 3 FREE R VALUE TEST SET COUNT : 2802
REMARK 3 ESTIMATED ERROR OF FREE R VALUE : 0.005
REMARK 3
REMARK 3 FIT IN THE HIGHEST RESOLUTION BIN.
REMARK 3 TOTAL NUMBER OF BINS USED : 6
REMARK 3 BIN RESOLUTION RANGE HIGH (A) : 2.00
REMARK 3 BIN RESOLUTION RANGE LOW (A) : 2.13
REMARK 3 BIN COMPLETENESS (WORKING+TEST) (%) : 97.90
REMARK 3 REFLECTIONS IN BIN (WORKING SET) : 8532
REMARK 3 BIN R VALUE (WORKING SET) : 0.2680
REMARK 3 BIN FREE R VALUE : 0.2910
REMARK 3 BIN FREE R VALUE TEST SET SIZE (%) : 5.00
REMARK 3 BIN FREE R VALUE TEST SET COUNT : 448
REMARK 3 ESTIMATED ERROR OF BIN FREE R VALUE : 0.014
REMARK 3
REMARK 3 NUMBER OF NON-HYDROGEN ATOMS USED IN REFINEMENT.
REMARK 3 PROTEIN ATOMS : 4276
REMARK 3 NUCLEIC ACID ATOMS : 0
REMARK 3 HETEROGEN ATOMS : 72
REMARK 3 SOLVENT ATOMS : 471
REMARK 3
REMARK 3 B VALUES.
REMARK 3 FROM WILSON PLOT (A**2) : 17.70
REMARK 3 MEAN B VALUE (OVERALL, A**2) : 35.50
REMARK 3 OVERALL ANISOTROPIC B VALUE.
REMARK 3 B11 (A**2) : 7.78000
REMARK 3 B22 (A**2) : -7.47000
REMARK 3 B33 (A**2) : -0.31000
REMARK 3 B12 (A**2) : 0.00000
REMARK 3 B13 (A**2) : 0.00000
REMARK 3 B23 (A**2) : 0.00000
REMARK 3
REMARK 3 ESTIMATED COORDINATE ERROR.
REMARK 3 ESD FROM LUZZATI PLOT (A) : 0.23
REMARK 3 ESD FROM SIGMAA (A) : 0.22
REMARK 3 LOW RESOLUTION CUTOFF (A) : 5.00
REMARK 3
REMARK 3 CROSS-VALIDATED ESTIMATED COORDINATE ERROR.
REMARK 3 ESD FROM C-V LUZZATI PLOT (A) : 0.28
REMARK 3 ESD FROM C-V SIGMAA (A) : 0.24
REMARK 3
REMARK 3 RMS DEVIATIONS FROM IDEAL VALUES.
REMARK 3 BOND LENGTHS (A) : 0.006
REMARK 3 BOND ANGLES (DEGREES) : 1.30
REMARK 3 DIHEDRAL ANGLES (DEGREES) : 21.40
REMARK 3 IMPROPER ANGLES (DEGREES) : 0.84
REMARK 3
REMARK 3 ISOTROPIC THERMAL MODEL : RESTRAINED
REMARK 3
REMARK 3 ISOTROPIC THERMAL FACTOR RESTRAINTS. RMS SIGMA
REMARK 3 MAIN-CHAIN BOND (A**2) : NULL ; NULL
REMARK 3 MAIN-CHAIN ANGLE (A**2) : NULL ; NULL
REMARK 3 SIDE-CHAIN BOND (A**2) : NULL ; NULL
REMARK 3 SIDE-CHAIN ANGLE (A**2) : NULL ; NULL
REMARK 3
REMARK 3 BULK SOLVENT MODELING.
REMARK 3 METHOD USED : FLAT MODEL
REMARK 3 KSOL : 0.35
REMARK 3 BSOL : 51.02
REMARK 3
REMARK 3 NCS MODEL : NULL
REMARK 3
REMARK 3 NCS RESTRAINTS. RMS SIGMA/WEIGHT
REMARK 3 GROUP 1 POSITIONAL (A) : NULL ; NULL
REMARK 3 GROUP 1 B-FACTOR (A**2) : NULL ; NULL
REMARK 3
REMARK 3 PARAMETER FILE 1 : PROTEIN_REP.PARAM
REMARK 3 PARAMETER FILE 2 : WATER_REP.PARAM
REMARK 3 PARAMETER FILE 3 : ION.PARAM
REMARK 3 PARAMETER FILE 4 : INH.PAR
REMARK 3 PARAMETER FILE 5 : NULL
REMARK 3 TOPOLOGY FILE 1 : PROTEIN.TOP
REMARK 3 TOPOLOGY FILE 2 : WATER.TOP
REMARK 3 TOPOLOGY FILE 3 : ION.TOP
REMARK 3 TOPOLOGY FILE 4 : INH.TOP
REMARK 3 TOPOLOGY FILE 5 : NULL
REMARK 3
REMARK 3 OTHER REFINEMENT REMARKS: NULL
REMARK 4
REMARK 4 1YVX COMPLIES WITH FORMAT V. 3.15, 01-DEC-08
REMARK 100
REMARK 100 THIS ENTRY HAS BEEN PROCESSED BY RCSB ON 28-FEB-05.
REMARK 100 THE RCSB ID CODE IS RCSB031979.
REMARK 200
REMARK 200 EXPERIMENTAL DETAILS
REMARK 200 EXPERIMENT TYPE : X-RAY DIFFRACTION
REMARK 200 DATE OF DATA COLLECTION : NULL
REMARK 200 TEMPERATURE (KELVIN) : 100
REMARK 200 PH : 6.0
REMARK 200 NUMBER OF CRYSTALS USED : 1
REMARK 200
REMARK 200 SYNCHROTRON (Y/N) : Y
REMARK 200 RADIATION SOURCE : ALS
REMARK 200 BEAMLINE : 8.3.1
REMARK 200 X-RAY GENERATOR MODEL : NULL
REMARK 200 MONOCHROMATIC OR LAUE (M/L) : M
REMARK 200 WAVELENGTH OR RANGE (A) : 1.100028
REMARK 200 MONOCHROMATOR : NULL
REMARK 200 OPTICS : NULL
REMARK 200
REMARK 200 DETECTOR TYPE : CCD
REMARK 200 DETECTOR MANUFACTURER : ADSC QUANTUM 210
REMARK 200 INTENSITY-INTEGRATION SOFTWARE : BLU-ICE
REMARK 200 DATA SCALING SOFTWARE : SCALEPACK
REMARK 200
REMARK 200 NUMBER OF UNIQUE REFLECTIONS : 55391
REMARK 200 RESOLUTION RANGE HIGH (A) : 2.000
REMARK 200 RESOLUTION RANGE LOW (A) : 40.000
REMARK 200 REJECTION CRITERIA (SIGMA(I)) : 0.000
REMARK 200
REMARK 200 OVERALL.
REMARK 200 COMPLETENESS FOR RANGE (%) : 99.3
REMARK 200 DATA REDUNDANCY : 3.500
REMARK 200 R MERGE (I) : NULL
REMARK 200 R SYM (I) : 0.07900
REMARK 200 <I/SIGMA(I)> FOR THE DATA SET : 9.8000
REMARK 200
REMARK 200 IN THE HIGHEST RESOLUTION SHELL.
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE HIGH (A) : 2.00
REMARK 200 HIGHEST RESOLUTION SHELL, RANGE LOW (A) : 2.07
REMARK 200 COMPLETENESS FOR SHELL (%) : NULL
REMARK 200 DATA REDUNDANCY IN SHELL : 3.30
REMARK 200 R MERGE FOR SHELL (I) : NULL
REMARK 200 R SYM FOR SHELL (I) : 0.49600
REMARK 200 <I/SIGMA(I)> FOR SHELL : 1.800
REMARK 200
REMARK 200 DIFFRACTION PROTOCOL: SINGLE WAVELENGTH
REMARK 200 METHOD USED TO DETERMINE THE STRUCTURE: MOLECULAR REPLACEMENT
REMARK 200 SOFTWARE USED: CNS
REMARK 200 STARTING MODEL: NULL
REMARK 200
REMARK 200 REMARK: NULL
REMARK 280
REMARK 280 CRYSTAL
REMARK 280 SOLVENT CONTENT, VS (%): 61.40
REMARK 280 MATTHEWS COEFFICIENT, VM (ANGSTROMS**3/DA): 3.10
REMARK 280
REMARK 280 CRYSTALLIZATION CONDITIONS: 15% PEG 8000, 0.2M AMMONIUM
REMARK 280 SULPHATE, 80MM SODIUM CITRATE PH 6.0, 7% GLYCEROL, 4% 1,6
REMARK 280 HEXANEDIOL AND 1% BENZAMIDINE, VAPOR DIFFUSION, HANGING DROP,
REMARK 280 TEMPERATURE 298K
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY
REMARK 290 SYMMETRY OPERATORS FOR SPACE GROUP: C 2 2 21
REMARK 290
REMARK 290 SYMOP SYMMETRY
REMARK 290 NNNMMM OPERATOR
REMARK 290 1555 X,Y,Z
REMARK 290 2555 -X,-Y,Z+1/2
REMARK 290 3555 -X,Y,-Z+1/2
REMARK 290 4555 X,-Y,-Z
REMARK 290 5555 X+1/2,Y+1/2,Z
REMARK 290 6555 -X+1/2,-Y+1/2,Z+1/2
REMARK 290 7555 -X+1/2,Y+1/2,-Z+1/2
REMARK 290 8555 X+1/2,-Y+1/2,-Z
REMARK 290
REMARK 290 WHERE NNN -> OPERATOR NUMBER
REMARK 290 MMM -> TRANSLATION VECTOR
REMARK 290
REMARK 290 CRYSTALLOGRAPHIC SYMMETRY TRANSFORMATIONS
REMARK 290 THE FOLLOWING TRANSFORMATIONS OPERATE ON THE ATOM/HETATM
REMARK 290 RECORDS IN THIS ENTRY TO PRODUCE CRYSTALLOGRAPHICALLY
REMARK 290 RELATED MOLECULES.
REMARK 290 SMTRY1 1 1.000000 0.000000 0.000000 0.00000
REMARK 290 SMTRY2 1 0.000000 1.000000 0.000000 0.00000
REMARK 290 SMTRY3 1 0.000000 0.000000 1.000000 0.00000
REMARK 290 SMTRY1 2 -1.000000 0.000000 0.000000 0.00000
REMARK 290 SMTRY2 2 0.000000 -1.000000 0.000000 0.00000
REMARK 290 SMTRY3 2 0.000000 0.000000 1.000000 62.20550
REMARK 290 SMTRY1 3 -1.000000 0.000000 0.000000 0.00000
REMARK 290 SMTRY2 3 0.000000 1.000000 0.000000 0.00000
REMARK 290 SMTRY3 3 0.000000 0.000000 -1.000000 62.20550
REMARK 290 SMTRY1 4 1.000000 0.000000 0.000000 0.00000
REMARK 290 SMTRY2 4 0.000000 -1.000000 0.000000 0.00000
REMARK 290 SMTRY3 4 0.000000 0.000000 -1.000000 0.00000
REMARK 290 SMTRY1 5 1.000000 0.000000 0.000000 30.55650
REMARK 290 SMTRY2 5 0.000000 1.000000 0.000000 107.40850
REMARK 290 SMTRY3 5 0.000000 0.000000 1.000000 0.00000
REMARK 290 SMTRY1 6 -1.000000 0.000000 0.000000 30.55650
REMARK 290 SMTRY2 6 0.000000 -1.000000 0.000000 107.40850
REMARK 290 SMTRY3 6 0.000000 0.000000 1.000000 62.20550
REMARK 290 SMTRY1 7 -1.000000 0.000000 0.000000 30.55650
REMARK 290 SMTRY2 7 0.000000 1.000000 0.000000 107.40850
REMARK 290 SMTRY3 7 0.000000 0.000000 -1.000000 62.20550
REMARK 290 SMTRY1 8 1.000000 0.000000 0.000000 30.55650
REMARK 290 SMTRY2 8 0.000000 -1.000000 0.000000 107.40850
REMARK 290 SMTRY3 8 0.000000 0.000000 -1.000000 0.00000
REMARK 290
REMARK 290 REMARK: NULL
REMARK 300
REMARK 300 BIOMOLECULE: 1
REMARK 300 SEE REMARK 350 FOR THE AUTHOR PROVIDED AND/OR PROGRAM
REMARK 300 GENERATED ASSEMBLY INFORMATION FOR THE STRUCTURE IN
REMARK 300 THIS ENTRY. THE REMARK MAY ALSO PROVIDE INFORMATION ON
REMARK 300 BURIED SURFACE AREA.
REMARK 350
REMARK 350 COORDINATES FOR A COMPLETE MULTIMER REPRESENTING THE KNOWN
REMARK 350 BIOLOGICALLY SIGNIFICANT OLIGOMERIZATION STATE OF THE
REMARK 350 MOLECULE CAN BE GENERATED BY APPLYING BIOMT TRANSFORMATIONS
REMARK 350 GIVEN BELOW. BOTH NON-CRYSTALLOGRAPHIC AND
REMARK 350 CRYSTALLOGRAPHIC OPERATIONS ARE GIVEN.
REMARK 350
REMARK 350 BIOMOLECULE: 1
REMARK 350 AUTHOR DETERMINED BIOLOGICAL UNIT: MONOMERIC
REMARK 350 APPLY THE FOLLOWING TO CHAINS: A
REMARK 350 BIOMT1 1 1.000000 0.000000 0.000000 0.00000
REMARK 350 BIOMT2 1 0.000000 1.000000 0.000000 0.00000
REMARK 350 BIOMT3 1 0.000000 0.000000 1.000000 0.00000
REMARK 465
REMARK 465 MISSING RESIDUES
REMARK 465 THE FOLLOWING RESIDUES WERE NOT LOCATED IN THE
REMARK 465 EXPERIMENT. (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN
REMARK 465 IDENTIFIER; SSSEQ=SEQUENCE NUMBER; I=INSERTION CODE.)
REMARK 465
REMARK 465 M RES C SSSEQI
REMARK 465 SER A 549
REMARK 465 TRP A 550
REMARK 465 PHE A 551
REMARK 465 THR A 552
REMARK 465 VAL A 553
REMARK 465 GLY A 554
REMARK 465 ALA A 555
REMARK 465 GLY A 556
REMARK 465 GLY A 557
REMARK 465 GLY A 558
REMARK 465 ASP A 559
REMARK 465 ILE A 560
REMARK 465 TYR A 561
REMARK 465 HIS A 562
REMARK 465 SER A 563
REMARK 465 VAL A 564
REMARK 465 SER A 565
REMARK 465 ARG A 566
REMARK 465 ALA A 567
REMARK 465 ARG A 568
REMARK 465 PRO A 569
REMARK 465 ARG A 570
REMARK 500
REMARK 500 GEOMETRY AND STEREOCHEMISTRY
REMARK 500 SUBTOPIC: TORSION ANGLES
REMARK 500
REMARK 500 TORSION ANGLES OUTSIDE THE EXPECTED RAMACHANDRAN REGIONS:
REMARK 500 (M=MODEL NUMBER; RES=RESIDUE NAME; C=CHAIN IDENTIFIER;
REMARK 500 SSEQ=SEQUENCE NUMBER; I=INSERTION CODE).
REMARK 500
REMARK 500 STANDARD TABLE:
REMARK 500 FORMAT:(10X,I3,1X,A3,1X,A1,I4,A1,4X,F7.2,3X,F7.2)
REMARK 500
REMARK 500 EXPECTED VALUES: GJ KLEYWEGT AND TA JONES (1996). PHI/PSI-
REMARK 500 CHOLOGY: RAMACHANDRAN REVISITED. STRUCTURE 4, 1395 - 1400
REMARK 500
REMARK 500 M RES CSSEQI PSI PHI
REMARK 500 PRO A 22 -80.98 -69.05
REMARK 500 SER A 27 44.36 -75.33
REMARK 500 SER A 29 123.70 -170.98
REMARK 500 LEU A 30 -77.02 -19.24
REMARK 500 ASN A 35 75.26 -160.36
REMARK 500 LYS A 36 33.86 -98.85
REMARK 500 ALA A 97 125.20 -39.35
REMARK 500 LEU A 260 -56.83 -134.76
REMARK 500 ALA A 348 59.14 -152.53
REMARK 500 HIS A 402 85.69 -43.95
REMARK 500 VAL A 424 -64.82 -101.27
REMARK 500 THR A 476 60.74 61.57
REMARK 500 LEU A 536 97.96 61.41
REMARK 500 PRO A 540 50.12 -68.33
REMARK 500 ALA A 542 47.79 -75.75
REMARK 500 ARG A 543 18.89 -143.43
REMARK 500
REMARK 500 REMARK: NULL
REMARK 525
REMARK 525 SOLVENT
REMARK 525
REMARK 525 THE SOLVENT MOLECULES HAVE CHAIN IDENTIFIERS THAT
REMARK 525 INDICATE THE POLYMER CHAIN WITH WHICH THEY ARE MOST
REMARK 525 CLOSELY ASSOCIATED. THE REMARK LISTS ALL THE SOLVENT
REMARK 525 MOLECULES WHICH ARE MORE THAN 5A AWAY FROM THE
REMARK 525 NEAREST POLYMER CHAIN (M = MODEL NUMBER;
REMARK 525 RES=RESIDUE NAME; C=CHAIN IDENTIFIER; SSEQ=SEQUENCE
REMARK 525 NUMBER; I=INSERTION CODE):
REMARK 525
REMARK 525 M RES CSSEQI
REMARK 525 HOH A 929 DISTANCE = 6.63 ANGSTROMS
REMARK 800
REMARK 800 SITE
REMARK 800 SITE_IDENTIFIER: AC1
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 571
REMARK 800 SITE_IDENTIFIER: AC2
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 572
REMARK 800 SITE_IDENTIFIER: AC3
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 573
REMARK 800 SITE_IDENTIFIER: AC4
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 574
REMARK 800 SITE_IDENTIFIER: AC5
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 575
REMARK 800 SITE_IDENTIFIER: AC6
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 576
REMARK 800 SITE_IDENTIFIER: AC7
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 577
REMARK 800 SITE_IDENTIFIER: AC8
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 578
REMARK 800 SITE_IDENTIFIER: AC9
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE SO4 A 579
REMARK 800 SITE_IDENTIFIER: BC1
REMARK 800 EVIDENCE_CODE: SOFTWARE
REMARK 800 SITE_DESCRIPTION: BINDING SITE FOR RESIDUE IPC A 580
REMARK 900
REMARK 900 RELATED ENTRIES
REMARK 900 RELATED ID: 1YUY RELATED DB: PDB
REMARK 900 RELATED ID: 1YVZ RELATED DB: PDB
REMARK 999
REMARK 999 SEQUENCE AUTHOR STATES THAT A PCR FUSION APPROACH WAS USED, AS
REMARK 999 IT IS WIDELY USED TO CONSTRUCT GENES WHEN NO ACTUAL TEMPLATE IS
REMARK 999 AVAILABLE (EG BLIGHT JK ET AL. SCIENCE. 2000 DEC 8;290(5498):
REMARK 999 1972-4) TO BUILT A CONSENSUS CDNA OF THE HCV POLYMERASE GENOTYPE
REMARK 999 2A. THE CONSENSUS SEQUENCE WAS BASED ON THE ALIGNMENT OF AT
REMARK 999 LEAST TEN COMPLETE SEQUENCES AVAILABLE IN THE NCBI DATABANK. THE
REMARK 999 GENE WAS PRODUCED COULD BE CONSIDERED SYNTHETIC AS IT WAS BASED
REMARK 999 ON THESE APPROXIMATELY 10 POLYMERASE SEQUENCES (AND WILL
REMARK 999 THEREFORE NOT CORRESPOND TO ANY SPECIFIC POLYMERASE SEQUENCE
REMARK 999 FOUND IN NATURE) AND THE CODON USAGE WAS ALSO MODIFIED TO
REMARK 999 MAXIMIZE THE EXPRESSION IN BACTERIA.
DBREF 1YVX A 1 570 UNP P26660 POLG_HCVJ6 2443 3012
SEQADV 1YVX PRO A 25 UNP P26660 SER 2467 SEE REMARK 999
SEQADV 1YVX THR A 150 UNP P26660 ALA 2592 SEE REMARK 999
SEQADV 1YVX ALA A 156 UNP P26660 PRO 2598 SEE REMARK 999
SEQADV 1YVX LYS A 212 UNP P26660 ARG 2654 SEE REMARK 999
SEQADV 1YVX VAL A 309 UNP P26660 ILE 2751 SEE REMARK 999
SEQADV 1YVX PRO A 391 UNP P26660 SER 2833 SEE REMARK 999
SEQRES 1 A 570 SER MET SER TYR SER TRP THR GLY ALA LEU ILE THR PRO
SEQRES 2 A 570 CYS SER PRO GLU GLU GLU LYS LEU PRO ILE ASN PRO LEU
SEQRES 3 A 570 SER ASN SER LEU LEU ARG TYR HIS ASN LYS VAL TYR CYS
SEQRES 4 A 570 THR THR SER LYS SER ALA SER LEU ARG ALA LYS LYS VAL
SEQRES 5 A 570 THR PHE ASP ARG MET GLN VAL LEU ASP ALA TYR TYR ASP
SEQRES 6 A 570 SER VAL LEU LYS ASP ILE LYS LEU ALA ALA SER LYS VAL
SEQRES 7 A 570 SER ALA ARG LEU LEU THR LEU GLU GLU ALA CYS GLN LEU
SEQRES 8 A 570 THR PRO PRO HIS SER ALA ARG SER LYS TYR GLY PHE GLY
SEQRES 9 A 570 ALA LYS GLU VAL ARG SER LEU SER GLY ARG ALA VAL ASN
SEQRES 10 A 570 HIS ILE LYS SER VAL TRP LYS ASP LEU LEU GLU ASP SER
SEQRES 11 A 570 GLN THR PRO ILE PRO THR THR ILE MET ALA LYS ASN GLU
SEQRES 12 A 570 VAL PHE CYS VAL ASP PRO THR LYS GLY GLY LYS LYS ALA
SEQRES 13 A 570 ALA ARG LEU ILE VAL TYR PRO ASP LEU GLY VAL ARG VAL
SEQRES 14 A 570 CYS GLU LYS MET ALA LEU TYR ASP VAL THR GLN LYS LEU
SEQRES 15 A 570 PRO GLN ALA VAL MET GLY ALA SER TYR GLY PHE GLN TYR
SEQRES 16 A 570 SER PRO ALA GLN ARG VAL GLU PHE LEU LEU LYS ALA TRP
SEQRES 17 A 570 ALA GLU LYS LYS ASP PRO MET GLY PHE SER TYR ASP THR
SEQRES 18 A 570 ARG CYS PHE ASP SER THR VAL THR GLU ARG ASP ILE ARG
SEQRES 19 A 570 THR GLU GLU SER ILE TYR GLN ALA CYS SER LEU PRO GLU
SEQRES 20 A 570 GLU ALA ARG THR ALA ILE HIS SER LEU THR GLU ARG LEU
SEQRES 21 A 570 TYR VAL GLY GLY PRO MET PHE ASN SER LYS GLY GLN SER
SEQRES 22 A 570 CYS GLY TYR ARG ARG CYS ARG ALA SER GLY VAL LEU THR
SEQRES 23 A 570 THR SER MET GLY ASN THR ILE THR CYS TYR VAL LYS ALA
SEQRES 24 A 570 LEU ALA ALA CYS LYS ALA ALA GLY ILE VAL ALA PRO THR
SEQRES 25 A 570 MET LEU VAL CYS GLY ASP ASP LEU VAL VAL ILE SER GLU
SEQRES 26 A 570 SER GLN GLY THR GLU GLU ASP GLU ARG ASN LEU ARG ALA
SEQRES 27 A 570 PHE THR GLU ALA MET THR ARG TYR SER ALA PRO PRO GLY
SEQRES 28 A 570 ASP PRO PRO ARG PRO GLU TYR ASP LEU GLU LEU ILE THR
SEQRES 29 A 570 SER CYS SER SER ASN VAL SER VAL ALA LEU GLY PRO GLN
SEQRES 30 A 570 GLY ARG ARG ARG TYR TYR LEU THR ARG ASP PRO THR THR
SEQRES 31 A 570 PRO ILE ALA ARG ALA ALA TRP GLU THR VAL ARG HIS SER
SEQRES 32 A 570 PRO VAL ASN SER TRP LEU GLY ASN ILE ILE GLN TYR ALA
SEQRES 33 A 570 PRO THR ILE TRP VAL ARG MET VAL LEU MET THR HIS PHE
SEQRES 34 A 570 PHE SER ILE LEU MET ALA GLN ASP THR LEU ASP GLN ASN
SEQRES 35 A 570 LEU ASN PHE GLU MET TYR GLY SER VAL TYR SER VAL SER
SEQRES 36 A 570 PRO LEU ASP LEU PRO ALA ILE ILE GLU ARG LEU HIS GLY
SEQRES 37 A 570 LEU ASP ALA PHE SER LEU HIS THR TYR THR PRO HIS GLU
SEQRES 38 A 570 LEU THR ARG VAL ALA SER ALA LEU ARG LYS LEU GLY ALA
SEQRES 39 A 570 PRO PRO LEU ARG ALA TRP LYS SER ARG ALA ARG ALA VAL
SEQRES 40 A 570 ARG ALA SER LEU ILE SER ARG GLY GLY ARG ALA ALA VAL
SEQRES 41 A 570 CYS GLY ARG TYR LEU PHE ASN TRP ALA VAL LYS THR LYS
SEQRES 42 A 570 LEU LYS LEU THR PRO LEU PRO GLU ALA ARG LEU LEU ASP
SEQRES 43 A 570 LEU SER SER TRP PHE THR VAL GLY ALA GLY GLY GLY ASP
SEQRES 44 A 570 ILE TYR HIS SER VAL SER ARG ALA ARG PRO ARG
HET SO4 A 571 5
HET SO4 A 572 5
HET SO4 A 573 5
HET SO4 A 574 5
HET SO4 A 575 5
HET SO4 A 576 5
HET SO4 A 577 5
HET SO4 A 578 5
HET SO4 A 579 5
HET IPC A 580 27
HETNAM SO4 SULFATE ION
HETNAM IPC 3-[ISOPROPYL(4-METHYLBENZOYL)AMINO]-5-PHENYLTHIOPHENE-
HETNAM 2 IPC 2-CARBOXYLIC ACID
FORMUL 2 SO4 9(O4 S 2-)
FORMUL 11 IPC C22 H21 N O3 S
FORMUL 12 HOH *471(H2 O)
HELIX 1 1 THR A 41 LYS A 43 5 3
HELIX 2 2 SER A 44 THR A 53 1 10
HELIX 3 3 ASP A 61 SER A 76 1 16
HELIX 4 4 THR A 84 LEU A 91 1 8
HELIX 5 5 GLY A 104 SER A 110 1 7
HELIX 6 6 SER A 112 ASP A 129 1 18
HELIX 7 7 ASP A 164 GLY A 188 1 25
HELIX 8 8 ALA A 189 TYR A 195 5 7
HELIX 9 9 SER A 196 GLU A 210 1 15
HELIX 10 10 CYS A 223 VAL A 228 1 6
HELIX 11 11 THR A 229 ALA A 242 1 14
HELIX 12 12 PRO A 246 LEU A 260 1 15
HELIX 13 13 THR A 286 GLY A 307 1 22
HELIX 14 14 GLY A 328 TYR A 346 1 19
HELIX 15 15 ASP A 359 ILE A 363 5 5
HELIX 16 16 PRO A 388 GLU A 398 1 11
HELIX 17 17 SER A 407 TYR A 415 1 9
HELIX 18 18 THR A 418 VAL A 424 1 7
HELIX 19 19 VAL A 424 ASP A 437 1 14
HELIX 20 20 SER A 455 LEU A 457 5 3
HELIX 21 21 ASP A 458 GLY A 468 1 11
HELIX 22 22 LEU A 469 SER A 473 5 5
HELIX 23 23 THR A 478 GLY A 493 1 16
HELIX 24 24 PRO A 496 ARG A 514 1 19
HELIX 25 25 GLY A 515 PHE A 526 1 12
HELIX 26 26 ASN A 527 VAL A 530 5 4
SHEET 1 A 5 TYR A 4 TRP A 6 0
SHEET 2 A 5 SER A 273 ARG A 277 -1 O TYR A 276 N SER A 5
SHEET 3 A 5 GLY A 264 PHE A 267 -1 N MET A 266 O GLY A 275
SHEET 4 A 5 THR A 136 ALA A 140 1 N THR A 136 O PHE A 267
SHEET 5 A 5 LEU A 159 PRO A 163 -1 O ILE A 160 N MET A 139
SHEET 1 B 2 VAL A 37 CYS A 39 0
SHEET 2 B 2 VAL A 144 CYS A 146 -1 O PHE A 145 N TYR A 38
SHEET 1 C 3 PRO A 214 ASP A 220 0
SHEET 2 C 3 ASP A 319 GLU A 325 -1 O SER A 324 N MET A 215
SHEET 3 C 3 THR A 312 CYS A 316 -1 N CYS A 316 O ASP A 319
SHEET 1 D 2 ASN A 369 LEU A 374 0
SHEET 2 D 2 ARG A 380 THR A 385 -1 O ARG A 381 N ALA A 373
SHEET 1 E 2 LEU A 443 GLU A 446 0
SHEET 2 E 2 VAL A 451 VAL A 454 -1 O VAL A 454 N LEU A 443
SSBOND 1 CYS A 316 CYS A 366 1555 1555 2.03
SITE 1 AC1 7 PRO A 93 ARG A 168 LYS A 172 HOH A 732
SITE 2 AC1 7 HOH A 735 HOH A 776 HOH A 796
SITE 1 AC2 6 PRO A 94 GLY A 104 ALA A 105 LYS A 106
SITE 2 AC2 6 HOH A 731 HOH A 846
SITE 1 AC3 2 GLY A 516 ARG A 517
SITE 1 AC4 3 ARG A 505 LYS A 531 THR A 532
SITE 1 AC5 6 TRP A 6 ALA A 9 SER A 46 ALA A 49
SITE 2 AC5 6 HOH A 645 HOH A 802
SITE 1 AC6 3 GLU A 230 TYR A 276 ARG A 278
SITE 1 AC7 5 THR A 329 GLU A 330 GLN A 436 LEU A 482
SITE 2 AC7 5 IPC A 580
SITE 1 AC8 3 HIS A 34 ASN A 35 LYS A 36
SITE 1 AC9 6 SER A 367 ARG A 386 THR A 390 HOH A 694
SITE 2 AC9 6 HOH A 856 HOH A 981
SITE 1 BC1 12 ARG A 422 LEU A 474 HIS A 475 THR A 476
SITE 2 BC1 12 TYR A 477 LEU A 497 ARG A 498 TRP A 528
SITE 3 BC1 12 SO4 A 577 HOH A 745 HOH A 767 HOH A 888
CRYST1 61.113 214.817 124.411 90.00 90.00 90.00 C 2 2 21 8
ORIGX1 1.000000 0.000000 0.000000 0.00000
ORIGX2 0.000000 1.000000 0.000000 0.00000
ORIGX3 0.000000 0.000000 1.000000 0.00000
SCALE1 0.016363 0.000000 0.000000 0.00000
SCALE2 0.000000 0.004655 0.000000 0.00000
SCALE3 0.000000 0.000000 0.008038 0.00000
(ATOM LINES ARE NOT SHOWN.)
END
