Correction for “Genetic confirmation for a central role for TNFα
in the direct action of thyroid stimulating hormone on the skele-
ton,” by Li Sun, Ling-Ling Zhu, Ping Lu, Tony Yuen, Jianhua Li,
Risheng Ma, Ramkumari Baliram, Surinder S. Moonga, Peng Liu,
Alberta Zallone, Maria I. New, Terry F. Davies, and Mone Zaidi,
which appeared in issue 24, June 11, 2013, of Proc Natl Acad Sci
USA (110:9891–9896; first published May 28, 2013; 10.1073/
The authors note that the author name Ramkumari Baliram
should instead appear as Ramkumarie Baliram. The corrected
author line appears below. The online version has been corrected.
Li Sun, Ling-Ling Zhu, Ping Lu, Tony Yuen, Jianhua Li,
Risheng Ma, Ramkumarie Baliram, Surinder S. Moonga,
Peng Liu, Alberta Zallone, Maria I. New, Terry F. Davies,
and Mone Zaidi
Correction for “IKK epsilon kinase is crucial for viral G protein-
coupled receptor tumorigenesis,” by Yi Wang, Xiaolu Lu,
Lining Zhu, Yan Shen, Shylet Chengedza, Hao Feng, Laiyee Wang,
Jae U. Jung, Julio S. Gutkind, and Pinghui Feng, which appeared
in issue 27, July 2, 2013, of Proc Natl Acad Sci USA (110:11139–
11144; first published June 14, 2013; 10.1073/pnas.1219829110).
The authors note that the author name Julio S. Gutkind
should instead appear as J. Silvio Gutkind. The corrected author
line appears below. The online version has been corrected.
Yi Wang, Xiaolu Lu, Lining Zhu, Yan Shen,
Shylet Chengedza, Hao Feng, Laiyee Wang, Jae U. Jung,
J. Silvio Gutkind, and Pinghui Feng
Correction for “Beginning of viniculture in France,” by Patrick E.
McGovern, Benjamin P. Luley, Nuria Rovira, Armen Mirzoian,
Michael P. Callahan, Karen E. Smith, Gretchen R. Hall, Theodore
Davidson, and Joshua M. Henkin, which appeared in issue 25,
June 18, 2013, of Proc Natl Acad Sci USA (110:10147–10152; first
published June 3, 2013; 10.1073/pnas.1216126110).
The authors note that on page 10151, left column, fourth full
paragraph, lines 8–11, “However, such exploitation and the
morphological transition between wild and domestic grapes is
not attested until at least the third century B.C., particularly at
Port Ariane, about a half kilometer distant from Lattara (26)”
should instead appear as “However, such exploitation and the
morphological transition between wild and domestic grapes is
not attested until at least the seventh–sixth century B.C., par-
ticularly at Port Ariane, about a half kilometer distant from
Correction for “Elasto-inertial turbulence,” by Devranjan Samanta,
Yves Dubief, Markus Holzner, Christof Schäfer, Alexander N.
Morozov, Christian Wagner, and Björn Hof, which appeared in
issue 26, June 25, 2013, of Proc Natl Acad Sci USA (110:10557–
10562; first published June 11, 2013; 10.1073/pnas.1219666110).
The authors note that the following statement should be added
to the Acknowledgments: “Y.D. gratefully acknowledges the
Vermont Advanced Computing Core, supported by NASA (NNX-
08AO96G), which provided the computational resources.”
| July 23, 2013
| vol. 110
| no. 30www.pnas.org
linked ubiquitin chain-mediated activation of the IKK complex,
consistingofIKKγ andIKKα and/orIKKβ (17).TheinducibleIKKe
was originally implicated in oncogenesis of breast cancer by an
siRNA screen, via inducing NF-κB activation (28). Accumulating
studies indicate that elevated IKKe expression is consistently ob-
served in multiple forms of malignant tumors (29), such as prostate
and ovarian cancers. However, the mechanism by which NF-κB is
activated by IKKe remains largely unknown. We found that IKKe,
when activated by kGPCR, phosphorylated RelA at Ser468 and
promoted its nuclear translocation, agreeing with the findings that
RelA Ser468p up-regulates the expression of a subset of inflam-
matory genes (32). Interestingly, previous reports, including our
findings from virus-infected cells (37), showed that the Ser468
phosphorylation primes RelA for degradation (38, 39). By contrast,
kGPCR expressionactivates IKKe that phosphorylates andactivates
RelA. These observations suggest that additional signaling events,
downstream of kGPCR, may switch RelA toward NF-κB activation.
The IKK-related IKKe was originally discovered as being in-
ducible by inflammatory cytokines (26). The involvement of IKKe
in the carcinogenesis of KS fits with the inflammatory and angio-
genic nature of KS tumors. The induction of IKKe expression
is more robust in human and mouse tumors than in kGPCR-
expressing endothelial cells, suggesting that in vivo paracrine
mechanisms of immune activation of IKKe are crucial and domi-
it is a general belief that induction of IKKe kinase activity largely
stems from an up-regulation of protein expression. The molecular
mechanisms governing IKKe activation at the posttranslational
level, in analogy to IKKβ activation, remain unknown. IKKe is
highly expressed in immunecells, including B and T lymphoid cells
(26). Our observation that IKKe is very abundant in tumor cells of
mouse KS-like lesions suggests that nonimmune cells, such as en-
distribution of IKKe is remarkably diverse in tumor cells, ranging
from being plasma membrane-associated, cytosolic punctate, and
nuclear. Similar patterns have been reported previously (28). High
levels of IKKe were also observed in human KS tumors, although
is predominantly expressed during lytic infection, KS tumor tissues
are mainly composed of endothelial cells that are latently infected
with KSHV. Conceivably, latent gene products may alter IKKe to
influence KSHV infection and diseases thereof in vivo. Indeed, our
results indicate that Kaposin B and, to a lesser extent, vCyclin, ac-
tivate IKKe, but not NF-κB. Conversely, vFLIP activates NF-κB,
butnot IKKe. These observations suggest that latentgene products
may function together to instigate IKKe and NF-κB activation. In
sum, ourstudy identifies IKKe as a key molecule linking kGPCR to
NF-κB activation and tumorigenesis. The potent inhibition of the
kinase-dead IKKeK38A on kGPCR tumorigenesis suggests that
IKKe represents a potential drug target to treat human KS.
Materials and Methods
Information on reagents and experimental procedures is given in SI Materials
and Methods. Included topics are animal experiments, immunoblotting,
qRT-PCR, immunofluorescence microscopy, EMSA, and H&E. All statistical
analyses were done with unpaired, two-tailed Student’s t test.
ACKNOWLEDGMENTS. We thank Ms. Yuqi Wang and Lisa Arneson for
assistance with the maintenance of mouse colonies and John Shelton and
Lillian Young for histology. This work was supported by National Cancer
Institute Grant R01 CA134241, National Institute of Health DE021445,
American Cancer Society Grant RSG-11-162-01-MPC (to P.F.), and National
Institute of Health U19:A1083025 (to J.U.J.).
1. Soulier J, et al. (1995) Kaposi’s sarcoma-associated herpesvirus-like DNA sequences in
multicentric Castleman’s disease. Blood 86(4):1276–1280.
2. Cesarman E, Chang Y, Moore PS, Said JW, Knowles DM (1995) Kaposi’s sarcoma-
associated herpesvirus-like DNA sequences in AIDS-related body-cavity-based lym-
phomas. N Engl J Med 332(18):1186–1191.
3. Chang Y, et al. (1994) Identification of herpesvirus-like DNA sequences in AIDS-
associated Kaposi’s sarcoma. Science 266(5192):1865–1869.
4. Nador RG, Cesarman E, Knowles DM, Said JW (1995) Herpes-like DNA sequences in
a body-cavity-based lymphoma in an HIV-negative patient. N Engl J Med 333(14):943.
5. Rezza G, et al. (1999) Human herpesvirus 8 seropositivity and risk of Kaposi’s sarcoma
and other acquired immunodeficiency syndrome-related diseases. J Natl Cancer Inst
6. Moore PS, Chang Y (2001) Molecular virology of Kaposi’s sarcoma-associated her-
pesvirus. Philos Trans R Soc Lond B Biol Sci 356(1408):499–516.
7. Montaner S, et al. (2003) Endothelial infection with KSHV genes in vivo reveals that
vGPCR initiates Kaposi’s sarcomagenesis and can promote the tumorigenic potential
of viral latent genes. Cancer Cell 3(1):23–36.
8. Sun R, et al. (1999) Kinetics of Kaposi’s sarcoma-associated herpesvirus gene expres-
sion. J Virol 73(3):2232–2242.
9. Yang TY, et al. (2000) Transgenic expression of the chemokine receptor encoded by
human herpesvirus 8 induces an angioproliferative disease resembling Kaposi’s sar-
coma. J Exp Med 191(3):445–454.
10. Wess J (1997) G-protein-coupled receptors: Molecular mechanisms involved in re-
ceptor activation and selectivity of G-protein recognition. FASEB J 11(5):346–354.
11. Pierce KL, Premont RT, Lefkowitz RJ (2002) Seven-transmembrane receptors. Nat Rev
Mol Cell Biol 3(9):639–650.
12. Arvanitakis L, Geras-Raaka E, Varma A, Gershengorn MC, Cesarman E (1997) Human
herpesvirus KSHV encodes a constitutively active G-protein-coupled receptor linked
to cell proliferation. Nature 385(6614):347–350.
13. Dorsam RT, Gutkind JS (2007) G-protein-coupled receptors and cancer. Nat Rev Cancer
14. Sodhi A, et al. (2006) The TSC2/mTOR pathway drives endothelial cell transformation
induced by the Kaposi’s sarcoma-associated herpesvirus G protein-coupled receptor.
Cancer Cell 10(2):133–143.
15. Martin D, et al. (2011) PI3Kγ mediates kaposi’s sarcoma-associated herpesvirus vGPCR-
induced sarcomagenesis. Cancer Cell 19(6):805–813.
16. Martin D, Galisteo R, Ji Y, Montaner S, Gutkind JS (2008) An NF-kappaB gene ex-
pression signature contributes to Kaposi’s sarcoma virus vGPCR-induced direct and
paracrine neoplasia. Oncogene 27(13):1844–1852.
17. Chen ZJ (2005) Ubiquitin signalling in the NF-kappaB pathway. Nat Cell Biol 7(8):758–765.
18. Häcker H, Karin M (2006) Regulation and function of IKK and IKK-related kinases.
Sci STKE 2006(357):re13.
19. Perkins ND (2007) Integrating cell-signalling pathways with NF-kappaB and IKK
function. Nat Rev Mol Cell Biol 8(1):49–62.
20. Hayden MS, Ghosh S (2008) Shared principles in NF-kappaB signaling. Cell 132(3):
21. Siebenlist U, Franzoso G, Brown K (1994) Structure, regulation and function of NF-
kappa B. Annu Rev Cell Biol 10:405–455.
22. Verma IM, Stevenson JK, Schwarz EM, Van Antwerp D, Miyamoto S (1995) Rel/NF-
kappa B/I kappa B family: Intimate tales of association and dissociation. Genes Dev
23. Hanahan D, Weinberg RA (2011) Hallmarks of cancer: The next generation. Cell
24. Sharma S, et al. (2003) Triggering the interferon antiviral response through an IKK-
related pathway. Science 300(5622):1148–1151.
25. Fitzgerald KA, et al. (2003) IKKepsilon and TBK1 are essential components of the IRF3
signaling pathway. Nat Immunol 4(5):491–496.
26. Shimada T, et al. (1999) IKK-i, a novel lipopolysaccharide-inducible kinase that is re-
lated to IkappaB kinases. Int Immunol 11(8):1357–1362.
27. Tenoever BR, et al. (2007) Multiple functions of the IKK-related kinase IKKepsilon in
interferon-mediated antiviral immunity. Science 315(5816):1274–1278.
28. Boehm JS, et al. (2007) Integrative genomic approaches identify IKBKE as a breast
cancer oncogene. Cell 129(6):1065–1079.
29. Clément JF, Meloche S, Servant MJ (2008) The IKK-related kinases: From innate im-
munity to oncogenesis. Cell Res 18(9):889–899.
30. Hayward GS (2003) Initiation of angiogenic Kaposi’s sarcoma lesions. Cancer Cell 3(1):1–3.
31. Cesarman E, Mesri EA, Gershengorn MC (2000) Viral G protein-coupled receptor and
Kaposi’s sarcoma: A model of paracrine neoplasia? J Exp Med 191(3):417–422.
32. Moreno R, Sobotzik JM, Schultz C, Schmitz ML (2010) Specification of the NF-kappaB
transcriptional response by p65 phosphorylation and TNF-induced nuclear trans-
location of IKK epsilon. Nucleic Acids Res 38(18):6029–6044.
33. Mattioli I, et al. (2006) Inducible phosphorylation of NF-kappa B p65 at serine 468 by
T cell costimulation is mediated by IKK epsilon. J Biol Chem 281(10):6175–6183.
34. Guasparri I, Keller SA, Cesarman E (2004) KSHV vFLIP is essential for the survival of
infected lymphoma cells. J Exp Med 199(7):993–1003.
35. Reilly SM, et al. (2013) An inhibitor of the protein kinases TBK1 and IKK-varepsilon
improves obesity-related metabolic dysfunctions in mice. Nat Med 19(3):313–321.
36. Ben-Neriah Y, Karin M (2011) Inflammation meets cancer, with NF-κB as the match-
maker. Nat Immunol 12(8):715–723.
37. Dong X, et al. (2012) Murine gammaherpesvirus 68 evades host cytokine production
via replication transactivator-induced RelA degradation. J Virol 86(4):1930–1941.
38. Mao X, et al. (2009) GCN5 is a required cofactor for a ubiquitin ligase that targets NF-
kappaB/RelA. Genes Dev 23(7):849–861.
39. Dong X, Feng P (2011) Murine gamma herpesvirus 68 hijacks MAVS and IKKβ to
abrogate NF-κB activation and antiviral cytokine production. PLoS Pathog 7(11):
| www.pnas.org/cgi/doi/10.1073/pnas.1219829110Wang et al.