An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish

Jun Zou, Diana Tran, Mai Baalbaki, Ling Fung Tang, Annie Poon, Angelo Pelonero, Erron W. Titus, Christiana Yuan, Chenxu Shi, Shruthi Patchava, Elizabeth Halper, Jasmine Garg, Irina Movsesyan, Chaoying Yin, Roland Wu, Lisa D Wilsbacher, Jiandong Liu, Ronald L. Hager, Shaun R. Coughlin, Martin JinekClive R. Pullinger, John P. Kane, Daniel O. Hart, Pui Yan Kwok, Rahul C. Deo*

*Corresponding author for this work

Research output: Contribution to journalArticle

37 Citations (Scopus)

Abstract

Truncating mutations in the giant sarcomeric protein Titin result in dilated cardiomyopathy and skeletal myopathy. The most severely affected dilated cardiomyopathy patients harbor Titin truncations in the C-terminal two-thirds of the protein, suggesting that mutation position might influence disease mechanism. Using CRISPR/Cas9 technology, we generated six zebrafish lines with Titin truncations in the N-terminal and C-terminal regions. Although all exons were constitutive, C-terminal mutations caused severe myopathy whereas Nterminal mutations demonstrated mild phenotypes. Surprisingly, neither mutation type acted as a dominant negative. Instead, we found a conserved internal promoter at the precise position where divergence in disease severity occurs, with the resulting protein product partially rescuing Nterminal truncations. In addition to its clinical implications, our work may shed light on a longstanding mystery regarding the architecture of the sarcomere.

Original languageEnglish (US)
Article numbere09406
JournaleLife
Volume4
Issue numberOCTOBER2015
DOIs
StatePublished - Oct 16 2015

Fingerprint

Connectin
Zebrafish
Mutation
Clustered Regularly Interspaced Short Palindromic Repeats
Dilated Cardiomyopathy
Muscular Diseases
Proteins
Ports and harbors
Exons
Sarcomeres
Technology
Phenotype

ASJC Scopus subject areas

  • Neuroscience(all)
  • Immunology and Microbiology(all)
  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Zou, J., Tran, D., Baalbaki, M., Tang, L. F., Poon, A., Pelonero, A., ... Deo, R. C. (2015). An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish. eLife, 4(OCTOBER2015), [e09406]. https://doi.org/10.7554/eLife.09406
Zou, Jun ; Tran, Diana ; Baalbaki, Mai ; Tang, Ling Fung ; Poon, Annie ; Pelonero, Angelo ; Titus, Erron W. ; Yuan, Christiana ; Shi, Chenxu ; Patchava, Shruthi ; Halper, Elizabeth ; Garg, Jasmine ; Movsesyan, Irina ; Yin, Chaoying ; Wu, Roland ; Wilsbacher, Lisa D ; Liu, Jiandong ; Hager, Ronald L. ; Coughlin, Shaun R. ; Jinek, Martin ; Pullinger, Clive R. ; Kane, John P. ; Hart, Daniel O. ; Kwok, Pui Yan ; Deo, Rahul C. / An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish. In: eLife. 2015 ; Vol. 4, No. OCTOBER2015.
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title = "An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish",
abstract = "Truncating mutations in the giant sarcomeric protein Titin result in dilated cardiomyopathy and skeletal myopathy. The most severely affected dilated cardiomyopathy patients harbor Titin truncations in the C-terminal two-thirds of the protein, suggesting that mutation position might influence disease mechanism. Using CRISPR/Cas9 technology, we generated six zebrafish lines with Titin truncations in the N-terminal and C-terminal regions. Although all exons were constitutive, C-terminal mutations caused severe myopathy whereas Nterminal mutations demonstrated mild phenotypes. Surprisingly, neither mutation type acted as a dominant negative. Instead, we found a conserved internal promoter at the precise position where divergence in disease severity occurs, with the resulting protein product partially rescuing Nterminal truncations. In addition to its clinical implications, our work may shed light on a longstanding mystery regarding the architecture of the sarcomere.",
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Zou, J, Tran, D, Baalbaki, M, Tang, LF, Poon, A, Pelonero, A, Titus, EW, Yuan, C, Shi, C, Patchava, S, Halper, E, Garg, J, Movsesyan, I, Yin, C, Wu, R, Wilsbacher, LD, Liu, J, Hager, RL, Coughlin, SR, Jinek, M, Pullinger, CR, Kane, JP, Hart, DO, Kwok, PY & Deo, RC 2015, 'An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish', eLife, vol. 4, no. OCTOBER2015, e09406. https://doi.org/10.7554/eLife.09406

An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish. / Zou, Jun; Tran, Diana; Baalbaki, Mai; Tang, Ling Fung; Poon, Annie; Pelonero, Angelo; Titus, Erron W.; Yuan, Christiana; Shi, Chenxu; Patchava, Shruthi; Halper, Elizabeth; Garg, Jasmine; Movsesyan, Irina; Yin, Chaoying; Wu, Roland; Wilsbacher, Lisa D; Liu, Jiandong; Hager, Ronald L.; Coughlin, Shaun R.; Jinek, Martin; Pullinger, Clive R.; Kane, John P.; Hart, Daniel O.; Kwok, Pui Yan; Deo, Rahul C.

In: eLife, Vol. 4, No. OCTOBER2015, e09406, 16.10.2015.

Research output: Contribution to journalArticle

TY - JOUR

T1 - An internal promoter underlies the difference in disease severity between N-and C-terminal truncation mutations of titin in zebrafish

AU - Zou, Jun

AU - Tran, Diana

AU - Baalbaki, Mai

AU - Tang, Ling Fung

AU - Poon, Annie

AU - Pelonero, Angelo

AU - Titus, Erron W.

AU - Yuan, Christiana

AU - Shi, Chenxu

AU - Patchava, Shruthi

AU - Halper, Elizabeth

AU - Garg, Jasmine

AU - Movsesyan, Irina

AU - Yin, Chaoying

AU - Wu, Roland

AU - Wilsbacher, Lisa D

AU - Liu, Jiandong

AU - Hager, Ronald L.

AU - Coughlin, Shaun R.

AU - Jinek, Martin

AU - Pullinger, Clive R.

AU - Kane, John P.

AU - Hart, Daniel O.

AU - Kwok, Pui Yan

AU - Deo, Rahul C.

PY - 2015/10/16

Y1 - 2015/10/16

N2 - Truncating mutations in the giant sarcomeric protein Titin result in dilated cardiomyopathy and skeletal myopathy. The most severely affected dilated cardiomyopathy patients harbor Titin truncations in the C-terminal two-thirds of the protein, suggesting that mutation position might influence disease mechanism. Using CRISPR/Cas9 technology, we generated six zebrafish lines with Titin truncations in the N-terminal and C-terminal regions. Although all exons were constitutive, C-terminal mutations caused severe myopathy whereas Nterminal mutations demonstrated mild phenotypes. Surprisingly, neither mutation type acted as a dominant negative. Instead, we found a conserved internal promoter at the precise position where divergence in disease severity occurs, with the resulting protein product partially rescuing Nterminal truncations. In addition to its clinical implications, our work may shed light on a longstanding mystery regarding the architecture of the sarcomere.

AB - Truncating mutations in the giant sarcomeric protein Titin result in dilated cardiomyopathy and skeletal myopathy. The most severely affected dilated cardiomyopathy patients harbor Titin truncations in the C-terminal two-thirds of the protein, suggesting that mutation position might influence disease mechanism. Using CRISPR/Cas9 technology, we generated six zebrafish lines with Titin truncations in the N-terminal and C-terminal regions. Although all exons were constitutive, C-terminal mutations caused severe myopathy whereas Nterminal mutations demonstrated mild phenotypes. Surprisingly, neither mutation type acted as a dominant negative. Instead, we found a conserved internal promoter at the precise position where divergence in disease severity occurs, with the resulting protein product partially rescuing Nterminal truncations. In addition to its clinical implications, our work may shed light on a longstanding mystery regarding the architecture of the sarcomere.

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