that connect T-tubules and the SR membrane ( 8 ).
Here, we discovered that JP2 contains additional
regulatory domains that extend beyond its role as
a structural protein. An NLS in the N-terminal
region of JP2 is necessary for nuclear import of
the calpain-generated JP2NT truncate. Thus,
under stress conditions, calpain-mediated cleav-
age of JP2 serves two purposes: (i) It impairs the
bridging of T-tubules with the SR membrane
[contributing to cardiomyocyte ultrastructural
remodeling and E-C uncoupling ( 23 )], and (ii) it
liberates JP2NT, allowing JP2NT to translocate
to the nucleus and mediate transcriptional re-
programming. In addition, we found that the
a-helix region of JP2 contains a previously un-
appreciated DNA binding domain that medi-
ates selective binding to canonical TATA box
motifs and MRE. This DNA binding domain is
evolutionarily conserved, suggestive of a dual
function for JP2 as a structural protein and tran-
scriptional regulator in other species.
The development and progression of heart
failure involves diverse cellular and molecular
mechanisms ( 41 , 42 ). Our ChIP-seq and tran-
scriptomic profiling data suggest that JP2NT
suppresses gene transcription by targeting multi-
ple signaling pathways such as inflammatory re-
sponses, fibrosis, myocyte hypotrophy, and cell
death among others. Taken with the protective
effect of JP2NT overexpression in the setting of
cardiac stress, this study indicates that JP2NT is
an endogenous self-protective stress transducer
that conveys the E-C uncoupling signal to the nu-
cleus, regulates transcriptional reprogramming,
and ultimately attenuates the progression of heart
failure. As JP2 is abundant in all muscle cells
(cardiac, skeletal, and smooth muscle), JP2NT
may serve as a general protective mechanism
antagonizing stress-induced pathological remod-
eling related to many diseases.
REFERENCES AND NOTES
- D. E. Clapham, Calcium signaling.Cell 131 , 1047–1058 (2007).
doi:10.1016/j.cell.2007.11.028; pmid: 18083096 - D. M. Bers, Cardiac excitation-contraction coupling.Nature
415 , 198–205 (2002). doi:10.1038/415198a; pmid: 11805843 - H. Cheng, W. J. Lederer, M. B. Cannell, Calcium sparks:
Elementary events underlying excitation-contraction coupling
in heart muscle.Science 262 , 740–744 (1993). doi:10.1126/
science.8235594; pmid: 8235594 - M. B. Cannell, H. Cheng, W. J. Lederer, The control of calcium
release in heart muscle.Science 268 , 1045–1049 (1995).
doi:10.1126/science.7754384; pmid: 7754384 - J. R. López-López, P. S. Shacklock, C. W. Balke, W. G. Wier,
Local calcium transients triggered by single L-type calcium
channel currents in cardiac cells.Science 268 , 1042– 1045
(1995). doi:10.1126/science.7754383; pmid: 7754383 - S. Q. Wang, L. S. Song, E. G. Lakatta, H. Cheng, Ca2+signalling
between single L-type Ca2+channels and ryanodine receptors
in heart cells.Nature 410 , 592–596 (2001). doi:10.1038/
35069083 ; pmid: 11279498 - E. Page, M. Surdyk-Droske, Distribution, surface density, and
membrane area of diadic junctional contacts between plasma
membrane and terminal cisterns in mammalian ventricle.
Circ. Res. 45 , 260–267 (1979). doi:10.1161/01.RES.45.2.260;
pmid: 376173 - H. Takeshima, S. Komazaki, M. Nishi, M. Iino, K. Kangawa,
Junctophilins: A novel family of junctional membrane complex
proteins.Mol. Cell 6 ,11–22 (2000). pmid: 10949023 - M. Nishi, A. Mizushima, Ki. Nakagawara, H. Takeshima,
Characterization of human junctophilin subtype genes.
Biochem. Biophys. Res. Commun. 273 , 920–927 (2000).
doi:10.1006/bbrc.2000.3011; pmid: 10891348- R. J. van Oortet al., Disrupted junctional membrane complexes
and hyperactive ryanodine receptors after acute junctophilin
knockdown in mice.Circulation 123 , 979–988 (2011).
doi:10.1161/CIRCULATIONAHA.110.006437; pmid: 21339484 - A. Guoet al., Overexpression of junctophilin-2 does not enhance
baseline function but attenuates heart failure development after
cardiac stress.Proc. Natl. Acad. Sci. U.S.A. 111 ,12240– 12245
(2014). doi:10.1073/pnas.1412729111; pmid: 25092313 - A. M. Gómezet al., Defective excitation-contraction coupling in
experimental cardiac hypertrophy and heart failure.Science
276 , 800–806 (1997). doi:10.1126/science.276.5313.800;
pmid: 9115206 - S. E. Litwin, D. Zhang, J. H. Bridge, Dyssynchronous Ca2+sparks
in myocytes from infarcted hearts.Circ. Res. 87 ,1040– 1047
(2000). doi:10.1161/01.RES.87.11.1040;pmid: 11090550 - L. S. Songet al., Orphaned ryanodine receptors in the failing
heart.Proc. Natl. Acad. Sci. U.S.A. 103 , 4305–4310 (2006).
doi:10.1073/pnas.0509324103; pmid: 16537526 - M. Xuet al., Intermolecular failure of L-type Ca2+channel and
ryanodine receptor signaling in hypertrophy.PLOS Biol. 5 , e21
(2007). doi:10.1371/journal.pbio.0050021; pmid: 17214508 - A. Guo, C. Zhang, S. Wei, B. Chen, L. S. Song, Emerging
mechanisms of T-tubule remodelling in heart failure.
Cardiovasc. Res. 98 , 204–215 (2013). doi:10.1093/cvr/cvt020;
pmid: 23393229 - S. Weiet al., T-tubule remodeling during transition from
hypertrophy to heart failure.Circ. Res. 107 , 520–531 (2010).
doi:10.1161/CIRCRESAHA.109.212324; pmid: 20576937 - M. Xuet al., Mir-24 regulates junctophilin-2 expression in
cardiomyocytes.Circ. Res. 111 , 837–841 (2012). doi:10.1161/
CIRCRESAHA.112.277418; pmid: 22891046 - H. D. Wuet al., Ultrastructural remodelling of Ca2+signalling
apparatus in failing heart cells.Cardiovasc. Res. 95 , 430– 438
(2012). doi:10.1093/cvr/cvs195; pmid: 22707157 - M. Jianget al., JPH-2 interacts with Cai-handling proteins and
ion channels in dyads: Contribution to premature ventricular
contraction-induced cardiomyopathy.Heart Rhythm 13 ,743– 752
(2016). doi:10.1016/j.hrthm.2015.10.037; pmid: 26538326 - S. Minamisawaet al., Junctophilin type 2 is associated with
caveolin-3 and is down-regulated in the hypertrophic and
dilated cardiomyopathies.Biochem. Biophys. Res. Commun.
325 , 852–856 (2004). doi:10.1016/j.bbrc.2004.10.107;
pmid: 15541368 - H. B. Zhanget al., Ultrastructural uncoupling between
T-tubules and sarcoplasmic reticulum in human heart failure.
Cardiovasc. Res. 98 , 269–276 (2013). doi:10.1093/cvr/
cvt030; pmid: 23405000 - C. Y. Wuet al., Calpain-dependent cleavage of junctophilin-2
and T-tubule remodeling in a mouse model of reversible heart
failure.J. Am. Heart Assoc. 3 , e000527 (2014). doi:10.1161/
JAHA.113.000527; pmid: 24958777 - A. Guoet al., Molecular Determinants of Calpain-dependent
Cleavage of Junctophilin-2 Protein in Cardiomyocytes.
J. Biol. Chem. 290 , 17946–17955 (2015). doi:10.1074/
jbc.M115.652396; pmid: 26063807 - Y. Wanget al., Targeting Calpain for Heart Failure Therapy:
Implications From Multiple Murine Models.JACC Basic Transl.
Sci. 3 , 503–517 (2018). doi:10.1016/j.jacbts.2018.05.004;
pmid: 30175274 - J. D. Molkentinet al., A calcineurin-dependent transcriptional
pathway for cardiac hypertrophy.Cell 93 , 215–228 (1998).
doi:10.1016/S0092-8674(00)81573-1; pmid: 9568714 - N. Frey, T. A. McKinsey, E. N. Olson, Decoding calcium signals
involved in cardiac growth and function.Nat. Med. 6 ,
1221 – 1227 (2000). doi:10.1038/81321; pmid: 11062532 - R. Passieret al .,CaM kinase signaling induces cardiac
hypertrophy and activates the MEF2 transcription factor
in vivo.J. Clin. Invest. 105 , 1395–1406 (2000). doi:10.1172/
JCI8551; pmid: 10811847 - J. Backs, K. Song, S. Bezprozvannaya, S. Chang, E. N. Olson,
CaM kinase II selectively signals to histone deacetylase 4
during cardiomyocyte hypertrophy.J. Clin. Invest. 116 ,
1853 – 1864 (2006). doi:10.1172/JCI27438; pmid: 16767219 - X. Wuet al., Local InsP3-dependent perinuclear Ca2+
signaling in cardiac myocyte excitation-transcription coupling.
J. Clin. Invest. 116 , 675–682 (2006). doi:10.1172/JCI27374;
pmid: 16511602 - M. Colellaet al., Ca2+oscillation frequency decoding in cardiac
cell hypertrophy: Role of calcineurin/NFAT as Ca2+signal
integrators.Proc. Natl. Acad. Sci. U.S.A. 105 , 2859– 2864
(2008). doi:10.1073/pnas.0712316105; pmid: 18287024- S. R. Houser, J. D. Molkentin, Does contractile Ca2+control
calcineurin-NFAT signaling and pathological hypertrophy in
cardiac myocytes?Sci. Signal. 1 , pe31 (2008). doi:10.1126/
scisignal.125pe31; pmid: 18577756 - A. S. Galvezet al., Cardiomyocyte degeneration with calpain
deficiency reveals a critical role in protein homeostasis.
Circ. Res. 100 , 1071–1078 (2007). doi:10.1161/
01.RES.0000261938.28365.11; pmid: 17332428 - C. Patterson, A. L. Portbury, J. C. Schisler, M. S. Willis, Tear me
down: Role of calpain in the development of cardiac ventricular
hypertrophy.Circ. Res. 109 , 453–462 (2011). doi:10.1161/
CIRCRESAHA.110.239749; pmid: 21817165 - D. J. Williams, H. L. Puhl 3rd, S. R. Ikeda, Rapid modification of
proteins using a rapamycin-inducible tobacco etch virus
protease system.PLOS ONE 4 , e7474 (2009). doi:10.1371/
journal.pone.0007474; pmid: 19830250 - C. J. Sigristet al., PROSITE: A documented database
using patterns and profiles as motif descriptors.Brief. Bioinform.
3 ,265–274 (2002). doi:10.1093/bib/3.3.265;pmid: 12230035 - G. Narasimhanet al., Mining protein sequences for motifs.
J. Comput. Biol. 9 , 707–720 (2002). doi:10.1089/
106652702761034145 ; pmid: 12487759 - J. Wysocka, P. T. Reilly, W. Herr, Loss of HCF-1-chromatin
association precedes temperature-induced growth arrest of
tsBN67 cells.Mol. Cell. Biol. 21 , 3820–3829 (2001).
doi:10.1128/MCB.21.11.3820-3829.2001; pmid: 11340173 - D. Sayed, M. He, Z. Yang, L. Lin, M. Abdellatif, Transcriptional
regulation patterns revealed by high resolution chromatin
immunoprecipitation during cardiac hypertrophy.J. Biol. Chem.
288 , 2546–2558 (2013). doi:10.1074/jbc.M112.429449;
pmid: 23229551 - F. J. Naya, C. Wu, J. A. Richardson, P. Overbeek, E. N. Olson,
Transcriptional activity of MEF2 during mouse embryogenesis
monitored with a MEF2-dependent transgene.Development
126 , 2045–2052 (1999). pmid: 10207130 - J. O. Mudd, D. A. Kass, Tackling heart failure in the twenty-first
century.Nature 451 , 919–928 (2008). doi:10.1038/
nature06798; pmid: 18288181 - J. H. van Berlo, M. Maillet, J. D. Molkentin, Signaling effectors
underlying pathologic growth and remodeling of the heart.
J. Clin. Invest. 123 ,37–45 (2013). doi:10.1172/JCI62839;
pmid: 23281408
ACKNOWLEDGMENTS
We thank M. J. Welsh, K. P. Campbell, E. D. Abel, B. London,
L. Yang (University of Iowa), and S. R. W. Chen (University of
Calgary) for reading the manuscript and providing constructive
comments, S. R. Ikeda (NIAAA/NIH) for providing TEVp plasmids,
and W. Kutschke for technique aids in animal surgery.Funding:
This work was funded by NIH R01 HL090905, HL130346, VA
1I01BX002334 (L.S.S.), AHA 16SDG30820003 (A.G.); NIH R01
HL125436 (C.G.), OD019941 (B.W.), and China NSF 81570293 (J.H.).
Author contributions:L.S.S. supervised the project; A.G. and
L.S.S. designed the study; A.G., Y.W., B.C., J.Y., L.Y.Z., D.H., J.W.,
Y.S., Q.Z., C.C., R.W., and X.Z. performed the experiments and
data analysis; Y.W. (Yunhao) and A.G. performed bioinformatics
analyses; C.G., M.E.A, F.Z., K.F.A., C.M., M.P., W.Z., and J.H.
participated in supervision of experiments, data analysis,
interpretations and revision of the manuscript. A.G. and L.S.S.
wrote the manuscript. All authors reviewed the results and edited
and approved the final version of the manuscript.Competing
interests:The authors have no conflicting financial interests.Data
and materials availability:Microarray and Sequencing data are
available at NCBI Gene Expression Omnibus (GEO) with accession
numbers GSE121545, GSE121546, and GSE121547. All other data
are available in the manuscript or the supplementary materials.
SUPPLEMENTARY MATERIALS
http://www.sciencemag.org/content/362/6421/eaan3303/suppl/DC1
Materials and Methods
Figs. S1 to S10
Tables S1 to S5
References ( 43 – 55 )
8 April 2017; resubmitted 10 May 2018
Accepted 24 October 2018
Published online 8 November 2018
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