|
|
|
|
|
|
[1]
|
NUCLEOTIDE SEQUENCE [GENOMIC RNA].
PubMed=10640553 [NCBI, ExPASy, EBI, Israel, Japan]
Netolitzky D.J.,
Schmaltz F.L.,
Parker M.D.,
Rayner G.A.,
Fisher G.R.,
Trent D.W.,
Bader D.E.,
Nagata L.P.;
"Complete genomic RNA sequence of western equine encephalitis virus and expression of the structural genes.";
J. Gen. Virol. 81:151-159(2000).
|
[2]
|
NUCLEOTIDE SEQUENCE [GENOMIC RNA] OF 2430-2466.
STRAIN=BFS1703;
PubMed=3413072 [NCBI, ExPASy, EBI, Israel, Japan]
Hahn C.S.,
Lustig S.,
Strauss E.G.,
Strauss J.H.;
"Western equine encephalitis virus is a recombinant virus.";
Proc. Natl. Acad. Sci. U.S.A. 85:5997-6001(1988).
|
|
|
|
- FUNCTION: P123 and P123' are short-lived polyproteins, accumulating during early stage of infection. P123 is directly translated from the genome, whereas P123' is a product of the cleavage of P1234. They localize the viral replication complex to the cytoplasmic surface of modified endosomes and lysosomes. By interacting with nsP4, they start viral genome replication into antigenome. After these early events, P123 and P123' are cleaved sequentially into nsP1, nsP2 and nsP3/nsP3'. This sequence of delayed processing would allow correct assembly and membrane association of the RNA polymerase complex (By similarity).
- FUNCTION: nsP1 is a cytoplasmic capping enzyme. This function is necessary since all viral RNAs are synthesized in the cytoplasm, and host capping enzymes are restricted to the nucleus. The enzymatic reaction involves a covalent link between 7-methyl-GMP and nsP1, whereas eukaryotic capping enzymes form a covalent complex only with GMP. nsP1 capping would consist in the following reactions: GTP is first methylated and then forms the m7GMp-nsP1 complex, from which 7-methyl-GMP complex is transferred to the mRNA to create the cap structure. Palmitoylated nsP1 is remodeling host cell cytoskeleton, and induces filopodium-like structure formation at the surface of the host cell (By similarity).
- FUNCTION: nsP2 has two separate domain with different biological activities. The N-terminal section is part of the RNA polymerase complex and has RNA trisphosphatase and RNA helicase activity. The C-terminal section harbors a protease that specifically cleaves and releases the four mature proteins (By similarity).
- FUNCTION: nsP3 and nsP3' are essential for minus strand and subgenomic 26S mRNA synthesis (By similarity).
- FUNCTION: nsP4 is a RNA dependent RNA polymerase. It replicates genomic and antigenomic RNA by recognizing replications specific signals. Transcribes also a 26S subgenomic mRNA by initiating RNA synthesis internally on antigenomic RNA. This 26S mRNA encodes for structural proteins. nsP4 is a short-lived protein regulated by several ways: the opal codon readthrough and degradation by ubiquitin pathway (By similarity).
- CATALYTIC ACTIVITY: S-adenosyl-L-methionine + GTP = m7GTP.
- CATALYTIC ACTIVITY: m7GTP + (5')pp-Pur-mRNA = diphosphate + m7G(5')ppp-Pur-mRNA.
- CATALYTIC ACTIVITY: (5')ppp-mRNA + H2O = (5')pp-mRNA + phosphate.
- CATALYTIC ACTIVITY: A 5'-phosphopolynucleotide + H2O = a polynucleotide + phosphate.
- CATALYTIC ACTIVITY: NTP + H2O = NDP + phosphate.
- CATALYTIC ACTIVITY: Nucleoside triphosphate + RNA(n) = diphosphate + RNA(n+1).
- SUBUNIT: P123 interacts with nsP4; nsP1, nsP2, nsP3 and nsP4 interact with each other, and with uncharacterized host factors (By similarity).
- SUBCELLULAR LOCATION: Non-structural polyprotein: Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Note=Located on the cytoplasmic surface of modified endosomes and lysosomes, also called cytopathic vacuoles type I (CPVI). These vacuoles contain numerous small circular invaginations (spherules) which may be the sites of RNA synthesis.
- SUBCELLULAR LOCATION: P123: Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity).
- SUBCELLULAR LOCATION: P123': Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity).
- SUBCELLULAR LOCATION: mRNA-capping enzyme nsP1: Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Cell membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Cell projection, filopodium (By similarity). Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then a fraction of nsP1 localizes to the inner surface of the plasma membrane and its filopodial extensions (By similarity).
- SUBCELLULAR LOCATION: Protease/triphosphatase/NTPase/helicase nsP2: Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Nucleus (By similarity). Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then approximately half of nsP2 is found in the nucleus (By similarity).
- SUBCELLULAR LOCATION: Non-structural protein 3: Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Cytoplasm (By similarity). Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then nsP3 and nsP3' seems to aggregate in cytoplasm (By similarity).
- SUBCELLULAR LOCATION: Non-structural protein 3': Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Cytoplasm (By similarity). Note=In the late phase of infection, the polyprotein is quickly cleaved before localization to cellular membranes. Then nsP3 and nsP3' seems to aggregate in cytoplasm (By similarity).
- SUBCELLULAR LOCATION: RNA-directed RNA polymerase nsP4: Endosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity). Lysosome membrane; Peripheral membrane protein; Cytoplasmic side (By similarity).
- INDUCTION: Viral replication produces dsRNA in the late phase of infection, resulting in a strong activation of host EIF2AK2/PKR, leading to almost complete phosphorylation of EIF2A. This inactivates completely cellular translation initiation, resulting in a dramatic shutoff of proteins synthesis. Translation of viral non-structural polyprotein and all cellular proteins are stopped in infected cell between 2 and 4 hours post infection. Only the 26S mRNA is still translated into viral structural proteins, presumably through a unique mechanism of enhancer element which counteract the translation inhibition mediated by EIF2A. By doing this, the virus uses the cellular defense for its own advantage: shutoff of cellular translation allows to produce big amounts of structural proteins needed for the virus to bud out of the doomed cell.
- PTM: Specific enzymatic cleavages in vivo yield mature proteins. The polyprotein is synthesized as P123, or P1234 by stop codon readthrough. These polyproteins are processed differently depending on the stage of infection. In early stages, P1234 is first cleaved in trans, through its nsP2 protease activity, releasing P123' and nsP4. P123/P123' and nsP4 start to replicate the viral genome into its antigenome. After these early events, nsP1 is cleaved in cis by nsP2 protease, releasing the P23/P23' polyprotein. Cleavage of nsP1 exposes an 'activator' at the N-terminus of P23/P23' which induces its cleavage into nsP2 and nsP3 by the viral protease. This sequence of delayed processing would allow correct assembly and membrane association of the RNA-polymerase complex. In the late stage of infection, the presence of free nsP2 in the cytoplasm cleaves P1234 quickly into P12 and P34, then into the four nsP (By similarity).
- PTM: nsP1 is palmitoylated by host (By similarity).
- PTM: nsP4 is ubiquitinated; targets the protein for rapid degradation via the ubiquitin system (By similarity).
- MISCELLANEOUS: The genome encodes for P123, but readthrough of a terminator codon UGA occurs between the codons for Asn-1852 and Arg-1853. This readthrough produces P1234, cleaved quickly by nsP2 into P123' and nsP4. Further processing of p123' gives nsP1, nsP2 and nsP3' which is 6 amino-acids longer than nsP3 since the cleavage site is after the readthrough. This unusual molecular mechanism ensures that few nsP4 are produced compared to other non-structural proteins. Mutant viruses with no alternative termination site grow significantly slower than wild-type virus.
- SIMILARITY: Contains 1 Macro domain.
- SIMILARITY: Contains 1 peptidase C9 domain [view classification].
- SIMILARITY: Contains 1 RdRp catalytic domain.
|
|
|
|
Copyrighted by the UniProt Consortium, see http://www.uniprot.org/terms.
Distributed under the Creative Commons Attribution-NoDerivs License.
|
|
|
|
|
|
|
|
|
|
|
|
|
|
|
| Length: 2466 AA [This is the length of the unprocessed precursor] |
Molecular weight: 274991 Da [This is the MW of the unprocessed precursor] |
CRC64: 646CB1A22FB63289 [This is a checksum on the sequence] |
|
10 20 30 40 50 60
MERIHVDLDA DSPYVKSLQR TFPQFEIEAR QVTDNDHANA RAFSHVATKL IESEVDRDQV
70 80 90 100 110 120
ILDIGSAPVR HAHSNHRYHC ICPMISAEDP DRLQRYAERL KKSDITDKNI ASKAADLLEV
130 140 150 160 170 180
MSTPDAETPS LCMHTDATCR YFGSVAVYQD VYAVHAPTSI YHQALKGVRT IYWIGFDTTP
190 200 210 220 230 240
FMYKNMAGSY PTYNTNWADE RVLEARNIGL GNSDLQESRL GKLSILRKKR LQPTNKIIFS
250 260 270 280 290 300
VGSTIYTEDR SLLRSWHLPN VFHLKGKSNF TGRCGTIVSC EGYVIKKITI SPGLYGKVEN
310 320 330 340 350 360
LASTMHREGF LSCKVTDTLR GERVSFAVCT YVPATLCDQM TGILATDVSV DDAQKLLVGL
370 380 390 400 410 420
NQRIVVNGRT QRNTNTMQNY LLPVVAQAFS RWAREHRADL DDEKELGVRE RTLTMGCCWA
430 440 450 460 470 480
FKTQKITSIY KKPGTQTIKK VPAVFDSFVI PRLTSHGLDM GFRRRLKLLL EPTVKPAPAI
490 500 510 520 530 540
TMADVEHLRG LQQEAEEVAA AEEIREALPP LLPEIEKETV EAEVDLIMQE AGAGSVETPR
550 560 570 580 590 600
GHIRVTSYPG EEKIGSYAIL SPQAVLNSEK LACIHPLAEQ VLVMTHKGRA GRYKVEPYHG
610 620 630 640 650 660
KVIVPEGTAV PVQDFQALSE SATIVFNERE FVNRYLHHIA INGGALNTDE EYYKTVKTQD
670 680 690 700 710 720
TDSEYVFDID ARKCVKREDA GPLCLTGDLV DPPFHEFAYE SLKTRPAAPH KVPTIGVYGV
730 740 750 760 770 780
PGSGKSGIIK SAVTKKDLVV SAKKENCAEI IRDVRRMRRM DVAARTVDSV LLNGVKHPVN
790 800 810 820 830 840
TLYIDEAFAC HAGTLLALIA IVKPKKVVLC GDPKQCGFFN MMCLKVHFNH DICTEVYHKS
850 860 870 880 890 900
ISRRCTQTVT AIVSTLFYDK RMKTVNPCAD KIIIDTTGTT KPHKDDLILT CFRGWVKQLQ
910 920 930 940 950 960
IDYKNHEIMT AAASQGLTRK GVYAVRYKVN ENPLYSQTSE HVNVLLTRTE KRIVWKTLAG
970 980 990 1000 1010 1020
DPWIKTLTAK YPGDFTASLD DWQREHDAIM ARVLDKPQTA DVFQNKVNVC WAKALEPVLA
1030 1040 1050 1060 1070 1080
TANIVLTRQQ WETLHPFKHD RAYSPEMALN FFCTRFFGVD LDSGLFSAPT VALTYRDQHW
1090 1100 1110 1120 1130 1140
DNSPGKNMYG LNREVAKELS RRYPCITKAV DTGRVADIRN NTIKDYSPTI NVVPLNRRLP
1150 1160 1170 1180 1190 1200
HSLIVDHKGQ GTTDHSGFLS KMKGKSVLVI GDPISIPGKK VESMGPLPTN TIRCDLDLGI
1210 1220 1230 1240 1250 1260
PSHVGKYDII FVNVRTPYRN HHYQQCEDHA IHHSMLTCKA VHHLNTGGTC VAIGYGLADR
1270 1280 1290 1300 1310 1320
ATENIITAVA RSFRFTRVCQ PKNTAENTEV LFVFFGKDNG NHTHDQDRLG VVLDNIYQGS
1330 1340 1350 1360 1370 1380
TRYEAGRAPA YRVIRGDISK SADQAIVNAA NSKGQPGSGV CGALYRKWPA AFDRQPIAVG
1390 1400 1410 1420 1430 1440
TARLVKHEPL IIHAVGPNFS KMPEPEGDLK LAAAYMSIAS IVNAERITKI SVPLLSTGIY
1450 1460 1470 1480 1490 1500
SGGKDRVMQS LHHLFTAFDT TDADVTIYCL DKQWETRIIE AIHRKESVEI LDDDKPVDID
1510 1520 1530 1540 1550 1560
LVRVHPNSSL AGRPGYSVNE GKLYSYLEGT RFHQTAKDIA EIHAMWPNKS EANEQICLYI
1570 1580 1590 1600 1610 1620
LGESMSSIRS KCPVEESEAS APPHTLPCLC NYAMTAERVY RLRSAKKEQF AVCSSFLLPK
1630 1640 1650 1660 1670 1680
YRITGVQKLQ CSKPVLFSGV VPPAVHPRKY AEIILETPPP PATTTVICEP TVPERIPSPV
1690 1700 1710 1720 1730 1740
ISRAPSAESL LSLGGVSFSS SATRSSTAWS DYDRRFVVTA DVHQANTSTW SIPSAPGLDV
1750 1760 1770 1780 1790 1800
QLPSDVTDSH WSIPSASGFE VRTPSVQDLT AECAKPRGLA EIMQDFNTAP FQFLSDYRPV
1810 1820 1830 1840 1850 1860
PAPRRRPIPS PRSTASAPPV PKPRRTKYQQ PPGVARAISE AELDEYIRQH SNRYEAGAYI
1870 1880 1890 1900 1910 1920
FSSETGQGHL QQKSVRQCKL QEPILDRAVH EKYYAPRLDL EREKMLQKKL QLCASEGNRS
1930 1940 1950 1960 1970 1980
RYQSRKVENM KAITAERLIS GLGTYLSSEV NPVECYRVNY PVPIYSSTVI NRFTSAEVAV
1990 2000 2010 2020 2030 2040
KTCNLVIQEN YPTVASYCIT DEYDAYLDMV DGASCCLDTA TFCPAKLRSY PKKHSYLQPE
2050 2060 2070 2080 2090 2100
IRSAVPSPIQ NTLQNVLAAA TKRNCNVTQM RELPVLDSAA FNVDCFKKYA CNDEYWDTFR
2110 2120 2130 2140 2150 2160
DNPIRLTTEN VTQYVTKLKG PKAAALFANT HNLKPLQEIP MDQFVMDLKR DVKVTPGTKH
2170 2180 2190 2200 2210 2220
TEERPKVQVI QAADPLATAY LCGIHRELVR RLNAVLLPNI HTLFDMSAED FDAIIAEHFH
2230 2240 2250 2260 2270 2280
HGDPVLETDI ASFDKSEDDA IAISALMILE DLGVDQPLLD LIEAAFGNIT SVHLPTGTRF
2290 2300 2310 2320 2330 2340
KFGAMMKSGM FLTLFVNTLV NIMIASRVLR ERLTTSACAA SIGDDNIVHG VVSDTLMAER
2350 2360 2370 2380 2390 2400
CATWLNMEVK IIDAVIGIKA PYFCGGFILV DQITGTACRV ADPLKRLFKL GKPLPVDDTQ
2410 2420 2430 2440 2450 2460
DCDRRRALHD EAMRWNRIGI TDELVKAVES RYEIILAGLI ITSLSTLAES VKNFKSIRGS
PITLYG
|
P13896 in FASTA format |
|
ExPASy Home page
SIB Switzerland
