Supplementary methods
SUPPLEMENTARY INFORMATION Exploration of Panviral Proteome: High-Throughput Cloning and Functional Implications in Virus-host Interactions Xiaobo Yu1,3, Xiaofang Bian1,3, Andrea Throop1, Lusheng Song1, Lerys Del Moral1, Jin Park1, Catherine Seiler1, Michael Fiacco1, Jason Steel1, Preston Hunter1, Justin Saul1, Jie Wang1, Ji Qiu1, James M. Pipas2, Joshua LaBaer1* 1
The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona
State University, Tempe, AZ 85287, USA 2
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA.
3
These authors contributed equally to this work.
*
Correspondence should be addressed to J. L. ([email protected]).
1
Supplementary methods Western blot The plasmid DNA of HCMV expression clones was prepared using our HT DNA preparation procedure as previously described [29]. Their concentrations were normalized to 1 µg/µL using H2O. The expression of HCMV proteins in pANT7_cGST vector with a C-terminal GST tag was performed by adding 1µg cDNA plasmid into 10 µL 2-fold diluted human HeLa lysate-based cell-free expression system (Thermo scientific, Rockford, IL) with the incubation of 2 hours (hrs) at 30oC. The expressed proteins were then mixed with 10 µL 2×SDS loading buffer containing 10% 2mercaptoethanol, and loaded into a 4-15% Tris-Criterion™ Precast Gel (Bio-rad, Hercules, CA). The protein electrophoresis was performed at 200V for 35 minutes (min). The proteins in gel were transferred to a nitrocellulose membrane using Trans-Blot® SD Semi-Dry Transfer Cell (Bio-rad, Hercules, CA) for 1 hr at 20 V. After blocking with 5% milk for 1 hr, the membrane was incubated with 1:3000 dilution of mouse anti-GST antibody overnight. After washing with PBST, the membrane was incubated with 1:5000 dilution of HRP conjugated sheep anti-mouse IgG antibody (Jackson Immunotech, West Grove, PA) for 1 hr at room temperature. The detection was performed using Chemiluminescence with SuperSignal West Femto Luminol/Enhancer Solution (Pierce, Rockford, IL. The expression of proteins was visually inspected by the band showed at their expected sizes on nitro-cellular membrane.
In-gel fluorescence 2
The expression of rubella viral proteins containing a C-terminal Halo tag and coxsackievirus proteins containing an N-terminal Halo tag was examined using in-gel fluorescence. Protein expression of viral ORF clones with Halo tag was performed by mixing 1 µL 1µg/µL cDNA plasmid and 9 µL diluted human HeLa expression solution, and incubating at 30oC for 2 hrs. The detection was performed by adding 10 µL of Alexa 660 conjugated Halo-ligand (Promega, Madison, WI) (6 µM) and following incubation for 1hr at room temperature. After protein electrophoresis, the protein gel was rinsed three times with H2O and scanned with an Amersham Bioscience Typhoon 9400 variable mode imager at 635 nm. All image adjustments were performed using Adobe® Photoshop® CS4 software (Adobe Systems, San Jose, CA).
Bead based pull-down assays Coupling of bait protein to the magnetic beads The Protein G coated Dynabeads (Life Technology, Carlsbad, CA) were completely re-suspended and transferred into a 1.5 mL centrifuge tube. The magnetic beads were separated from the solution using a magnet stand. After removal of the supernatant, the coupling of Dynabeads was performed by incubation with 100 µg/mL rabbit antiGST polyclonal antibody (GE Healthcare Biosciences, Piscataway, NJ) at room temperature for 2 hrs on a Eppendorf Thermomixer® R mixer incubator (Eppendorf, Hauppauge, NY) at 1400 rpm. After the reaction, the beads were washed three times with PBST and stored at 4oC. In parallel, the bait proteins were expressed by using 50 ng/µL plasmid cDNA in human cell-free expression system with the incubation of 2 hrs 3
at 30oC. The bait proteins were purified using anti-GST antibody coupled magnetic beads incubated for 2 hrs at 4oC.
Pull-down assays The Halo tagged rubella viral proteins were prepared by the same means as described above. To perform pull-down assay, the supernatant of the purified bait proteins coated magnetic beads was removed with the magnet. The resulting bait protein-beads were incubated with 25 µL of the C-terminal Halo tagged rubella viral proteins overnight at 4oC in the Eppendorf Thermomixer® R mixer incubator at 1400 rpm. After washing three times with PPI wash buffer, the detection was performed by adding 25 µL of Alex660 labeled Halo-ligand (6 µM) and incubating for 2 hrs at 4oC. Lastly, the resulting beads were washed three times with PBST and the supernatant was removed with the magnet. To dissociate the query and bait proteins from the beads, 20 µL 1×SDS loading buffer containing 10% 2-mercaptoethanol was added to the beads, and incubated for 5 min at 95°C. The dissociated proteins were ran on a 4-15% Tris-Criterion™ Precast Gel (Bio-rad, Hercules, CA) at 200 V for 35 min. After rinsing three times with diH2O, the protein gels were scanned in an Amersham Bioscience Typhoon 9400 variable mode imager at 635 nm. All image adjustments were performed using Adobe® Photoshop® CS4 software (Adobe Systems, San Jose, CA). Subsequent to in-gel fluorescence scanning, the bait protein with C-terminal GST tag in the same gel was examined with
4
western blot using mouse anti-GST antibody and HRP conjugated sheep anti-mouse IgG antibody.
5
Supplementary figures
Input of bead-based pull-down assays 209
Halo E1
Halo E2
P90
150
Halo Capsid
Halo P150
P150N
100 75
P150C
37 25
Figure S1 The input of Halo and rubella viral proteins used for the bead-based pulldown assays. Each protein has a C-terminal Halo tag and was detected using Alexa660 labeled Halo ligand.
6
Subcellular location
Cell membrane
15%
5%
37%
3% 2% 5%
Cytoplasm Nucleus Golgi
19%
Endoplasmic reticulum 14%
Mitochondrion Secreted. NA
Figure S2 The subcellular location of host target candidates for rubella virus. The annotation was performed using UniProt (Universal Protein Resource) database (www.uniprot.org/).
7
Coxsackievirus protein expression VP0
VP4
VP3
VP1
2A
2B
2C
3A
3C
RP
209
100 54 41 27 20
Figure S3 Examination of coxsackievirus protein expression using in-gel fluorescence. Each protein has an N-terminal Halo tag and was detected using Alexa660 labeled Halo ligand.
8
Supplementary Tables Table S1 Visualized inspection of HCMV ORFeome expression using western blot analysis. No.
Gene
Class
Protein
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41
UL104 UL46 UL48A UL80 UL80.5 UL85 UL86 UL49 UL76 UL79 UL92 UL95 RL10 UL100 UL115 UL128 UL130 UL131A UL132 UL33 UL37 UL4 UL55 UL73 UL74 UL74A UL75 UL78 US27 US28 RL11 RL13 UL1 UL10 UL11 UL119 UL120 UL121 UL124 UL133 UL136
capsid capsid capsid capsid capsid capsid capsid core core core core core envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane
capsid portal protein capsid triplex subunit 1 small capsid protein capsid maturation protease capsid scaffold protein capsid triplex subunit 2 major capsid protein protein UL49 nuclear protein UL24 protein UL79 protein UL92 protein UL95 envelope glycoprotein RL10 envelope glycoprotein M envelope glycoprotein L envelope protein UL128 envelope glycoprotein UL130 envelope protein UL131A envelope glycoprotein UL132 envelope glycoprotein UL33 envelope glycoprotein UL37 envelope glycoprotein UL4 envelope glycoprotein B envelope glycoprotein N envelope glycoprotein O envelope glycoprotein 24 envelope glycoprotein H envelope protein UL78 envelope glycoprotein US27 envelope protein US28 membrane glycoprotein RL11 membrane protein RL13 membrane protein UL1 membrane protein UL10 membrane glycoprotein UL11 membrane glycoprotein UL119 membrane protein UL120 membrane protein UL121 membrane protein UL124 protein UL133 protein UL136 9
Visualized inspection No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90
UL138 UL139 UL14 UL140 UL141 UL142 UL144 UL147A UL148 UL148A UL148B UL148C UL148D UL15A UL16 UL16 UL18 UL2 UL20 UL40 UL41A UL42 UL5 UL50 UL6 UL7 UL8 UL9 US10 US11 US12 US13 US14 US15 US16 US17 US18 US19 US2 US20 US21 US29 US3 US30 US34A US6 US7 US8 US9
membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane
protein UL138 membrane glycoprotein UL139 membrane protein UL14 protein UL140 membrane glycoprotein UL141 membrane glycoprotein UL142 membrane glycoprotein UL144 membrane protein UL147A membrane protein UL148 protein UL148A protein UL148B protein UL148C protein UL148D protein UL15A membrane glycoprotein UL16 membrane glycoprotein UL16 membrane glycoprotein UL18 protein UL2 membrane protein UL20 membrane glycoprotein UL40 protein UL41A protein UL42 protein UL5 nuclear egress membrane protein membrane protein UL6 membrane protein UL7 protein UL8 membrane glycoprotein UL9 membrane glycoprotein US10 membrane glycoprotein US11 membrane protein US12 membrane protein US13 membrane protein US14 membrane protein US15 membrane protein US16 membrane protein US17 membrane protein US18 membrane protein US19 membrane glycoprotein US2 membrane protein US20 membrane protein US21 protein US29 membrane glycoprotein US3 membrane protein US30 protein US34A membrane glycoprotein US6 membrane glycoprotein US7 membrane glycoprotein US8 membrane glycoprotein US9 10
Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139
RL1 RL5A RL6 UL102 UL105 UL112 UL114 UL117 UL122 UL123 UL145 UL154 UL17 UL19 UL21A UL27 UL29 UL30 UL31 UL34 UL38 UL44 UL51 UL52 UL53 UL54 UL56 UL57 UL69 UL70 UL72 UL84 UL87 UL89 UL91 UL98 US1 US23 US26 US31 US32 UL111A UL116 UL13 UL135 UL146 UL147 UL150 UL22A
other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other secreted secreted secreted secreted secreted secreted secreted secreted
protein RL1 protein RL5A protein RL6 helicase-primase subunit helicase-primase helicase subunit protein UL112 uracil-DNA glycosylase protein UL117 regulatory protein IE2 regulatory protein IE1 protein UL145 protein UL154 protein UL17 protein UL19 protein UL21A protein UL27 protein UL29 protein UL30 protein UL31 protein UL34 protein UL38 DNA polymerase processivity subunit DNA packaging protein UL33 DNA packaging protein UL32 nuclear egress lamina protein DNA polymerase catalytic subunit DNA packaging terminase subunit 2 single-stranded DNA-binding protein multifunctional expression regulator helicase-primase primase subunit deoxyuridine triphosphatase protein UL84 protein UL87 DNA packaging terminase subunit 1 protein UL91 deoxyribonuclease protein US1 protein US23 protein US26 protein US31 protein US32 interleukin-10 protein UL116 protein UL13 protein UL135 chemokine vCXCL1 chemokine vCXCL2 protein UL150 glycoprotein UL22A 11
Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No
140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165
US34 TRS1 UL103 IRS1 UL23 UL24 UL25 UL26 UL32 UL35 UL36 UL43 UL45 UL47 UL48 UL71 UL77 UL82 UL83 UL88 UL93 UL96 UL97 UL99 US22 US24
secreted tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument
protein US34 tegument protein TRS1 tegument protein UL7 tegument protein IRS1 tegument protein UL23 tegument protein UL24 tegument protein UL25 tegument protein UL26 tegument protein pp150 tegument protein UL35 tegument protein vICA tegument protein UL43 ribonucleotide reductase subunit 1 tegument protein UL37 large tegument protein tegument protein UL51 DNA packaging tegument protein UL25 tegument protein pp71 tegument protein pp65 tegument protein UL88 DNA packaging tegument protein UL17 tegument protein UL14 tegument serine/threonine protein kinase myristylated tegument protein tegument protein US22 tegument protein US24
12
Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
The Virginia G. Piper Center for Personalized Diagnostics, Biodesign Institute, Arizona
State University, Tempe, AZ 85287, USA 2
Department of Biological Sciences, University of Pittsburgh, Pennsylvania 15260, USA.
3
These authors contributed equally to this work.
*
Correspondence should be addressed to J. L. ([email protected]).
1
Supplementary methods Western blot The plasmid DNA of HCMV expression clones was prepared using our HT DNA preparation procedure as previously described [29]. Their concentrations were normalized to 1 µg/µL using H2O. The expression of HCMV proteins in pANT7_cGST vector with a C-terminal GST tag was performed by adding 1µg cDNA plasmid into 10 µL 2-fold diluted human HeLa lysate-based cell-free expression system (Thermo scientific, Rockford, IL) with the incubation of 2 hours (hrs) at 30oC. The expressed proteins were then mixed with 10 µL 2×SDS loading buffer containing 10% 2mercaptoethanol, and loaded into a 4-15% Tris-Criterion™ Precast Gel (Bio-rad, Hercules, CA). The protein electrophoresis was performed at 200V for 35 minutes (min). The proteins in gel were transferred to a nitrocellulose membrane using Trans-Blot® SD Semi-Dry Transfer Cell (Bio-rad, Hercules, CA) for 1 hr at 20 V. After blocking with 5% milk for 1 hr, the membrane was incubated with 1:3000 dilution of mouse anti-GST antibody overnight. After washing with PBST, the membrane was incubated with 1:5000 dilution of HRP conjugated sheep anti-mouse IgG antibody (Jackson Immunotech, West Grove, PA) for 1 hr at room temperature. The detection was performed using Chemiluminescence with SuperSignal West Femto Luminol/Enhancer Solution (Pierce, Rockford, IL. The expression of proteins was visually inspected by the band showed at their expected sizes on nitro-cellular membrane.
In-gel fluorescence 2
The expression of rubella viral proteins containing a C-terminal Halo tag and coxsackievirus proteins containing an N-terminal Halo tag was examined using in-gel fluorescence. Protein expression of viral ORF clones with Halo tag was performed by mixing 1 µL 1µg/µL cDNA plasmid and 9 µL diluted human HeLa expression solution, and incubating at 30oC for 2 hrs. The detection was performed by adding 10 µL of Alexa 660 conjugated Halo-ligand (Promega, Madison, WI) (6 µM) and following incubation for 1hr at room temperature. After protein electrophoresis, the protein gel was rinsed three times with H2O and scanned with an Amersham Bioscience Typhoon 9400 variable mode imager at 635 nm. All image adjustments were performed using Adobe® Photoshop® CS4 software (Adobe Systems, San Jose, CA).
Bead based pull-down assays Coupling of bait protein to the magnetic beads The Protein G coated Dynabeads (Life Technology, Carlsbad, CA) were completely re-suspended and transferred into a 1.5 mL centrifuge tube. The magnetic beads were separated from the solution using a magnet stand. After removal of the supernatant, the coupling of Dynabeads was performed by incubation with 100 µg/mL rabbit antiGST polyclonal antibody (GE Healthcare Biosciences, Piscataway, NJ) at room temperature for 2 hrs on a Eppendorf Thermomixer® R mixer incubator (Eppendorf, Hauppauge, NY) at 1400 rpm. After the reaction, the beads were washed three times with PBST and stored at 4oC. In parallel, the bait proteins were expressed by using 50 ng/µL plasmid cDNA in human cell-free expression system with the incubation of 2 hrs 3
at 30oC. The bait proteins were purified using anti-GST antibody coupled magnetic beads incubated for 2 hrs at 4oC.
Pull-down assays The Halo tagged rubella viral proteins were prepared by the same means as described above. To perform pull-down assay, the supernatant of the purified bait proteins coated magnetic beads was removed with the magnet. The resulting bait protein-beads were incubated with 25 µL of the C-terminal Halo tagged rubella viral proteins overnight at 4oC in the Eppendorf Thermomixer® R mixer incubator at 1400 rpm. After washing three times with PPI wash buffer, the detection was performed by adding 25 µL of Alex660 labeled Halo-ligand (6 µM) and incubating for 2 hrs at 4oC. Lastly, the resulting beads were washed three times with PBST and the supernatant was removed with the magnet. To dissociate the query and bait proteins from the beads, 20 µL 1×SDS loading buffer containing 10% 2-mercaptoethanol was added to the beads, and incubated for 5 min at 95°C. The dissociated proteins were ran on a 4-15% Tris-Criterion™ Precast Gel (Bio-rad, Hercules, CA) at 200 V for 35 min. After rinsing three times with diH2O, the protein gels were scanned in an Amersham Bioscience Typhoon 9400 variable mode imager at 635 nm. All image adjustments were performed using Adobe® Photoshop® CS4 software (Adobe Systems, San Jose, CA). Subsequent to in-gel fluorescence scanning, the bait protein with C-terminal GST tag in the same gel was examined with
4
western blot using mouse anti-GST antibody and HRP conjugated sheep anti-mouse IgG antibody.
5
Supplementary figures
Input of bead-based pull-down assays 209
Halo E1
Halo E2
P90
150
Halo Capsid
Halo P150
P150N
100 75
P150C
37 25
Figure S1 The input of Halo and rubella viral proteins used for the bead-based pulldown assays. Each protein has a C-terminal Halo tag and was detected using Alexa660 labeled Halo ligand.
6
Subcellular location
Cell membrane
15%
5%
37%
3% 2% 5%
Cytoplasm Nucleus Golgi
19%
Endoplasmic reticulum 14%
Mitochondrion Secreted. NA
Figure S2 The subcellular location of host target candidates for rubella virus. The annotation was performed using UniProt (Universal Protein Resource) database (www.uniprot.org/).
7
Coxsackievirus protein expression VP0
VP4
VP3
VP1
2A
2B
2C
3A
3C
RP
209
100 54 41 27 20
Figure S3 Examination of coxsackievirus protein expression using in-gel fluorescence. Each protein has an N-terminal Halo tag and was detected using Alexa660 labeled Halo ligand.
8
Supplementary Tables Table S1 Visualized inspection of HCMV ORFeome expression using western blot analysis. No.
Gene
Class
Protein
1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41
UL104 UL46 UL48A UL80 UL80.5 UL85 UL86 UL49 UL76 UL79 UL92 UL95 RL10 UL100 UL115 UL128 UL130 UL131A UL132 UL33 UL37 UL4 UL55 UL73 UL74 UL74A UL75 UL78 US27 US28 RL11 RL13 UL1 UL10 UL11 UL119 UL120 UL121 UL124 UL133 UL136
capsid capsid capsid capsid capsid capsid capsid core core core core core envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop envelop membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane
capsid portal protein capsid triplex subunit 1 small capsid protein capsid maturation protease capsid scaffold protein capsid triplex subunit 2 major capsid protein protein UL49 nuclear protein UL24 protein UL79 protein UL92 protein UL95 envelope glycoprotein RL10 envelope glycoprotein M envelope glycoprotein L envelope protein UL128 envelope glycoprotein UL130 envelope protein UL131A envelope glycoprotein UL132 envelope glycoprotein UL33 envelope glycoprotein UL37 envelope glycoprotein UL4 envelope glycoprotein B envelope glycoprotein N envelope glycoprotein O envelope glycoprotein 24 envelope glycoprotein H envelope protein UL78 envelope glycoprotein US27 envelope protein US28 membrane glycoprotein RL11 membrane protein RL13 membrane protein UL1 membrane protein UL10 membrane glycoprotein UL11 membrane glycoprotein UL119 membrane protein UL120 membrane protein UL121 membrane protein UL124 protein UL133 protein UL136 9
Visualized inspection No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 66 67 68 69 70 71 72 73 74 75 76 77 78 79 80 81 82 83 84 85 86 87 88 89 90
UL138 UL139 UL14 UL140 UL141 UL142 UL144 UL147A UL148 UL148A UL148B UL148C UL148D UL15A UL16 UL16 UL18 UL2 UL20 UL40 UL41A UL42 UL5 UL50 UL6 UL7 UL8 UL9 US10 US11 US12 US13 US14 US15 US16 US17 US18 US19 US2 US20 US21 US29 US3 US30 US34A US6 US7 US8 US9
membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane membrane
protein UL138 membrane glycoprotein UL139 membrane protein UL14 protein UL140 membrane glycoprotein UL141 membrane glycoprotein UL142 membrane glycoprotein UL144 membrane protein UL147A membrane protein UL148 protein UL148A protein UL148B protein UL148C protein UL148D protein UL15A membrane glycoprotein UL16 membrane glycoprotein UL16 membrane glycoprotein UL18 protein UL2 membrane protein UL20 membrane glycoprotein UL40 protein UL41A protein UL42 protein UL5 nuclear egress membrane protein membrane protein UL6 membrane protein UL7 protein UL8 membrane glycoprotein UL9 membrane glycoprotein US10 membrane glycoprotein US11 membrane protein US12 membrane protein US13 membrane protein US14 membrane protein US15 membrane protein US16 membrane protein US17 membrane protein US18 membrane protein US19 membrane glycoprotein US2 membrane protein US20 membrane protein US21 protein US29 membrane glycoprotein US3 membrane protein US30 protein US34A membrane glycoprotein US6 membrane glycoprotein US7 membrane glycoprotein US8 membrane glycoprotein US9 10
Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
91 92 93 94 95 96 97 98 99 100 101 102 103 104 105 106 107 108 109 110 111 112 113 114 115 116 117 118 119 120 121 122 123 124 125 126 127 128 129 130 131 132 133 134 135 136 137 138 139
RL1 RL5A RL6 UL102 UL105 UL112 UL114 UL117 UL122 UL123 UL145 UL154 UL17 UL19 UL21A UL27 UL29 UL30 UL31 UL34 UL38 UL44 UL51 UL52 UL53 UL54 UL56 UL57 UL69 UL70 UL72 UL84 UL87 UL89 UL91 UL98 US1 US23 US26 US31 US32 UL111A UL116 UL13 UL135 UL146 UL147 UL150 UL22A
other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other other secreted secreted secreted secreted secreted secreted secreted secreted
protein RL1 protein RL5A protein RL6 helicase-primase subunit helicase-primase helicase subunit protein UL112 uracil-DNA glycosylase protein UL117 regulatory protein IE2 regulatory protein IE1 protein UL145 protein UL154 protein UL17 protein UL19 protein UL21A protein UL27 protein UL29 protein UL30 protein UL31 protein UL34 protein UL38 DNA polymerase processivity subunit DNA packaging protein UL33 DNA packaging protein UL32 nuclear egress lamina protein DNA polymerase catalytic subunit DNA packaging terminase subunit 2 single-stranded DNA-binding protein multifunctional expression regulator helicase-primase primase subunit deoxyuridine triphosphatase protein UL84 protein UL87 DNA packaging terminase subunit 1 protein UL91 deoxyribonuclease protein US1 protein US23 protein US26 protein US31 protein US32 interleukin-10 protein UL116 protein UL13 protein UL135 chemokine vCXCL1 chemokine vCXCL2 protein UL150 glycoprotein UL22A 11
Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes No
140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 161 162 163 164 165
US34 TRS1 UL103 IRS1 UL23 UL24 UL25 UL26 UL32 UL35 UL36 UL43 UL45 UL47 UL48 UL71 UL77 UL82 UL83 UL88 UL93 UL96 UL97 UL99 US22 US24
secreted tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument tegument
protein US34 tegument protein TRS1 tegument protein UL7 tegument protein IRS1 tegument protein UL23 tegument protein UL24 tegument protein UL25 tegument protein UL26 tegument protein pp150 tegument protein UL35 tegument protein vICA tegument protein UL43 ribonucleotide reductase subunit 1 tegument protein UL37 large tegument protein tegument protein UL51 DNA packaging tegument protein UL25 tegument protein pp71 tegument protein pp65 tegument protein UL88 DNA packaging tegument protein UL17 tegument protein UL14 tegument serine/threonine protein kinase myristylated tegument protein tegument protein US22 tegument protein US24
12
Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes Yes
