Table 1.
UPR qPCR results for tunicamycin treated and VZV infected cells.
| Gene | Function | Group | TM treated | VZV infected | ||
|---|---|---|---|---|---|---|
| ΔΔCT | STD | ΔΔCT | STD | |||
| ARMET/MANF | ERSE-II regulated; reduces cell proliferation and UPR initiated apoptosis | ANTI | 5.0 | 0.2 | 6.1 | 0.5 |
| EDEM3 | ER degradation enhancer, mannosidase alpha-like 3 | ERAD | −1.5 | 0.9 | −4.7 | 0.9 |
| PPIA | Peptidylpropyl isomerase A (cyclophilin A) | ERAD | −1.4 | 0.2 | −0.7 | 0.5 |
| UBE2G2 | E2 ubiquitin conjugating enzyme G2 | ERAD | −0.9 | 0.2 | 0.4 | 0.5 |
| NPLOC4 (NPL4) | Regulates poly Ub on cytosolic side of ER membrane with VCP | ERAD | −0.7 | 0.2 | 1.9 | 0.5 |
| UBXN4/erasin | UBX domain protein 4—adaptor protein to VCP | ERAD | −0.5 | 0.3 | −6.1 | 0.5 |
| USP14 | Ubiquitin specific peptidase 14 in cytosol | ERAD | −0.4 | 0.3 | −0.3 | 0.5 |
| SEC62 | ERAD translocation pore formation | ERAD | −0.2 | 0.3 | −4.3 | 0.4 |
| UFD1L | Regulates poly Ub on cytosolic side of ER membrane with VCP | ERAD | 0.1 | 0.1 | 0.9 | 0.4 |
| UBE2J2 | E2 ubiquitin conjugating enzyme J2 | ERAD | 0.2 | 0.1 | −0.4 | 0.4 |
| ATXN3 | Ataxin 3—deubiquiting enzyme | ERAD | 0.8 | 0.2 | 3.8 | 0.5 |
| FBX06 | E3 ubiquition ligase of glycoproteins in ER lumen | ERAD | 0.9 | 0.1 | −2.1 | 0.4 |
| RNF5 | Ring Finger protein 5—E3 Ubiquitin ligase in ER membrane | ERAD | 1.1 | 0.1 | 1.1 | 0.4 |
| DERL1 | Derlin family member E3 Ubiquitin ligase in ER membrane | ERAD | 1.2 | 0.2 | 2.0 | 0.5 |
| VCP(p97) | Regulates poly Ub of translocated ER substrates with NPL4 and UFD1L | ERAD | 1.3 | 0.2 | −0.4 | 0.5 |
| EDEM1 | ER degradation enhancer, mannosidase alpha-like 1—trims mannose | ERAD | 1.4 | 0.2 | 1.5 | 0.4 |
| SEL1L | Adaptor protein of Derlin-3/HRD1 in ER membrane | ERAD | 2.0 | 0.3 | −2.6 | 0.4 |
| OS9 | Glycoprotein protein quality control | ERAD | 2.0 | 0.2 | −0.7 | 0.5 |
| DERL2 | Derlin family member, E3 Ubiquitin ligase in ER membrane | ERAD | 2.4 | 0.2 | −1.3 | 0.5 |
| SYVN1 (DER3/HRD1) | Synoviolin, Derlin family member E3 Ubiquitin ligase in ER membrane | ERAD | 2.7 | 0.1 | 1.9 | 0.4 |
| SELS | Selenoprotein S—oxidoreductase (oxidative stress) | ERAD | 3.0 | 0.2 | 1.6 | 0.5 |
| HERPUD1 (HERP) | Mediates degradation of ER Ca channels | ERAD | 4.3 | 0.3 | −1.9 | 0.5 |
| HSPA2 | HSP70 protein 2 | FOLD | −1.5 | 0.3 | 1.2 | 0.6 |
| HSPA4 | HSP70 protein 4 | FOLD | −0.8 | 0.3 | 1.4 | 0.5 |
| HSPA1B | HSP70 protein 1B | FOLD | −0.5 | 0.2 | 0.0 | 0.5 |
| GANAB (Glu II) | Glucosidase that trims N-linked glycans | FOLD | −0.5 | 0.3 | −1.6 | 0.5 |
| PRKCSH | Protein kinase C substrate 80K-H (subunit of glucosidase II) | FOLD | −0.4 | 0.2 | 1.7 | 0.5 |
| GANC | Glycosal hydoloysis | FOLD | −0.2 | 0.2 | 2.3 | 0.4 |
| TCP1 | Component of Chaperonin | FOLD | −0.2 | 0.1 | −0.7 | 0.4 |
| HSPA1L | HSP70 protein 1 like | FOLD | −0.1 | 0.2 | −3.0 | 0.4 |
| HSPA4L | HSP70 protein 4 like | FOLD | 0.0 | 0.3 | −1.7 | 0.5 |
| CCT4 | Chaperonin containing TCP1, subunit 4 (delta) | FOLD | 0.3 | 0.1 | −1.3 | 0.4 |
| CCT7 | Chaperonin containing TCP1, subunit 7 (eta) | FOLD | 0.4 | 0.2 | 0.4 | 0.5 |
| HSPH1 | HSP105 protein 1 | FOLD | 0.6 | 0.1 | 1.6 | 0.4 |
| PFDN5 | Prefoldin subunit 5; co-chaperone of Chaperonin complex | FOLD | 0.7 | 0.2 | 2.2 | 0.5 |
| PFDN2 | Prefoldin subunit 2 | FOLD | 1.0 | 0.2 | 1.5 | 0.5 |
| TOR1A | Torsion A—ATPase | FOLD | 1.0 | 0.2 | −1.0 | 0.5 |
| UGCGL2 (UGT2) | UDP-glucose ceramide glucosyltransferase-like 2 | FOLD | 1.0 | 0.2 | 1.6 | 0.5 |
| UGCGL1 (UGT1) | UDP-glucose ceramide glucosyltransferase-like 1 | FOLD | 1.1 | 0.2 | 0.9 | 0.5 |
| ERP44 | Thiol chaperone | FOLD | 1.5 | 0.3 | −1.8 | 0.5 |
| CALR | Calreticulin; glycoprotein folding chaperone | FOLD | 1.5 | 0.3 | −1.1 | 0.5 |
| RPN1 | Ribophorin 1—substrate specific facilitator of N-glycosylation | FOLD | 1.6 | 0.2 | −1.6 | 0.5 |
| ERO1L | Thiol oxidase governs redox state of ER (with Ca2+) | FOLD | 1.6 | 0.3 | 0.8 | 0.5 |
| DNAJC10 (ERdj5) | DNAJ (HSP40 homolog), subfamily C, member 10 | FOLD | 1.7 | 0.3 | −1.0 | 0.5 |
| DNAJB2 | DNAJ (HSP40 homolog), subfamily B, member 2 | FOLD | 1.8 | 0.2 | 2.0 | 0.5 |
| SEC63 | Regulates ER import of membrane proteins | FOLD | 2.1 | 0.2 | 0.3 | 0.4 |
| CANX | Calnexin; glycoprotein folding chaperone; binds Ca2+ | FOLD | 2.2 | 0.3 | 1.2 | 0.5 |
| SIL1(BAP) | Nucleotide exchange factor; binds BiP | FOLD | 2.4 | 0.1 | 2.0 | 0.4 |
| PDIA3 (ERP57) | Protein disulfide isomerase family A, member 3 | FOLD | 2.5 | 0.3 | 1.9 | 0.5 |
| DNAJC3 | DNAJ (HSP40 homolog), subfamily C, member 3 | FOLD | 2.6 | 0.1 | 2.4 | 0.4 |
| DNAJC4 | DNAJ (HSP40 homolog), subfamily C, member 4 | FOLD | 2.9 | 0.2 | 2.9 | 0.5 |
| ERO1LB | Thiol oxidase governs redox state of ER (with Ca2+) | FOLD | 3.8 | 0.2 | 1.5 | 0.4 |
| DNAJB9 (ERdj4) | DNAJ (HSP40 homolog), subfamily B, member 9 | FOLD | 5.5 | 0.2 | −2.9 | 0.5 |
| HSPA5 | HSP70 protein 5 GRP78 (BIP) | FOLD | 6.3 | 0.1 | 4.7 | 0.4 |
| SREBF2 | Sterol regulatory element binding TF 2 | LIPID | 0.9 | 0.1 | −1.9 | 0.4 |
| RNF139 (TRC8) | E3 Ubiquition ligase associated with INSIG | LIPID | −0.3 | 0.6 | −1.2 | 0.6 |
| INSIG2 | Insulin induced protein isoform 2; regulation of cholesterol synthesis | LIPID | 0.3 | 0.3 | 4.1 | 0.4 |
| INSIG1 | Insulin induced protein isoform 1; regulation of cholesterol synthesis | LIPID | 0.5 | 0.3 | 5.3 | 0.4 |
| AMFR (gp78) | Autocrine motility factor receptor; E3 Ub ligase; regulation of cholesterol | LIPID | 0.6 | 0.3 | 6.2 | 0.5 |
| SREBF1 | Sterol regulatory element binding TF 1 | LIPID | 1.1 | 0.2 | 0.3 | 0.5 |
| SCAP | Activates SREBF by cleaving it | LIPID | 1.6 | 0.2 | −0.3 | 0.5 |
| SERP1 (RAMP4) | Stress induced ER protein 1; ER salt channel regulation | LIPID | 2.7 | 0.3 | 2.9 | 0.6 |
| MAPK8 (JNK1) | Map kinase K8 aka JNK1; pro-apoptotic in response to TNFα | PRO | −0.8 | 0.1 | 2.2 | 0.4 |
| BAX | BCL2-associated X protein; induces release of COX-2 from mitochondria | PRO | 0.5 | 0.3 | −1.0 | 0.5 |
| MAPK9 (JNK2) | Mitogen-activated protein kinase 9 | PRO | 0.5 | 0.2 | 1.0 | 0.4 |
| HTRA2 | HTRA serine peptidase 4 | PRO | 0.6 | 0.2 | −0.2 | 0.5 |
| MAPK10 (JNK3) | Map kinase K10 aka JNK3; pro-apoptotic in neurons | PRO | 1.0 | 0.4 | 2.3 | 0.6 |
| HTRA4 | HTRA serine peptidase 2 | PRO | 1.4 | 0.4 | 4.0 | 0.6 |
| CHOP | Aka DDIT3/GADD153; ER stress associated apoptotic protein | PRO | 5.3 | 0.2 | 0.1 | 0.4 |
| MBTPS2/S2P | Membrane bound TF peptidase, site 2 (active in Golgi) | SENSOR | −0.2 | 0.3 | 1.0 | 0.4 |
| MBTPS1/S1P | Membrane bound TP peptidase, site 1 (active in Golgi; cleaves ATF6) | SENSOR | −0.1 | 0.2 | 0.1 | 0.4 |
| ERN1 (IRE1α) | IRE1α is an endonuclease that splices XBP1 upon activation | SENSOR | 0.0 | 0.2 | 1.6 | 0.4 |
| ATF6B | ATF6 beta | SENSOR | 0.5 | 0.3 | −3.0 | 0.5 |
| CREB3 (LUMAN) | OASIS (B-zip TF) family member; cell proliferation | SENSOR | 1.4 | 0.2 | 1.0 | 0.4 |
| ATF6 | Activating transcription factor 6 | SENSOR | 1.5 | 0.2 | 0.3 | 0.5 |
| EIF2AK3 (PERK) | ER stress sensor; PKR-like kinase | SENSOR | 1.6 | 0.2 | −4.2 | 0.5 |
| ERN2 (IRE1β) | ER to nucleus signaling protein 2 | SENSOR | 1.8 | 0.5 | 3.1 | 0.6 |
| NUCB1 | Nucleobindin 1; negative regulation of ATF6 | SENSOR | 3.1 | 0.2 | 4.1 | 0.5 |
| CREB3L3 (CREBH) | TF regulating lipogenesis and secretory pathway | SENSOR | 4.4 | 1.6 | 9.3 | 0.9 |
| XBP1 | X box binding protein 1; splicing by IRE1 activates XBP1 | TF | 0.7 | 0.1 | −0.4 | 0.4 |
| CEBPB (C/EBPβ) | Bzip TF with wide impact on cell cycle and proliferation | TF | 1.3 | 0.3 | −2.9 | 0.6 |
| ATF4 | Activates stress response (including CHOP) | TF | 1.7 | 0.2 | 1.5 | 0.5 |
| EIF2A | Eukaryotic translation initiation factor 2A | TRANS | 0.0 | 0.5 | 0.1 | 0.4 |
| PPP1R15A | Protein phosphatase 1, subunit 15A | TRANS | 1.8 | 0.2 | 1.1 | 0.4 |
Human fibroblast cells (MRC-5) were grown on glass coverslips in tissue culture plates then infected with VZV-32 infected MRC-5 cells or treated with tunicamycin (TM), a N-glycosylation inhibitor. At 72 hpi, RNA was extracted from the VZV-32 infected cultures. For the TM treated cultures, RNA was extracted at 24 h post-treatment. RNA from the VZV-32 infected, TM treated and uninfected cell cultures was then converted to cDNA which was applied to UPR specific PCR arrays (SA Biosciences) and real time PCR was carried out on an ABI 7000 PCR instrument The resulting CT values were then normalized (ΔCT) by housekeeping genes in the plate and then differences (ΔΔCT) between the uninfected and infected or tunicamycin treated values were computed and averaged. Abbreviations: anti-apoptotic (ANTI), ER associated degradation (ERAD), protein folding chaperones (FOLD), lipid and fat metabolism (LIPID), pro-apoptotic (PRO), ER stress sensor proteins (SENSOR), other transcription factors (TF) and protein translation associated proteins (TRANS). Error estimates correspond to standard deviation (STD).