Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2

James A.  Oo; Katalin  Pálfi; Timothy  Warwick; Ilka  Wittig; Cristian  Prieto-Garcia; Vigor  Matkovic; Ines  Tomašković; Frederike  Boos; Judit  Izquierdo Ponce; Tom  Teichmann; Kirill  Petriukov; Shaza  Haydar; Lars Maegdefessel; Zhiyuan  Wu; Minh  Duc Pham; Jaya  Krishnan; Andrew H Baker; Stefan  Günther; Helle D.  Ulrich; Ivan Dikic; Matthias S Leisegang; Ralf P Brandes

doi:10.1016/j.celrep.2022.111670

Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2

James A. Oo, Katalin Pálfi, Timothy Warwick, Ilka Wittig, Cristian Prieto-Garcia, Vigor Matkovic, Ines Tomašković, Frederike Boos, Judit Izquierdo Ponce, Tom Teichmann, Kirill Petriukov, Shaza Haydar, Lars Maegdefessel, Zhiyuan Wu, Minh Duc Pham, Jaya Krishnan, Andrew H Baker, Stefan Günther, Helle D. Ulrich, Ivan DikicMatthias S Leisegang, Ralf P Brandes^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract / Description of output

In healthy vessels, endothelial cells maintain a stable, differentiated and growth-arrested phenotype for years. Upon injury, a rapid phenotypic switch facilitates proliferation to restore tissue perfusion. Here we report the identification of the endothelial cell-enriched long non-coding RNA (lncRNA) PCAT19, which contributes to the proliferative switch and acts as a safeguard for the endothelial
genome. PCAT19 is enriched in confluent, quiescent endothelial cells and binds to the full replication protein A (RPA) complex in a DNA damage and cell cycle-related manner. Our results suggest that PCAT19 limits the phosphorylation of RPA2, primarily on the serine 33 (S33) residue, and thereby facilitates an appropriate DNA damage response while slowing cell cycle progression. Reduction in PCAT19 levels, either in response to loss of cell contacts or knockdown, promotes endothelial proliferation and angiogenesis. Collectively, PCAT19 acts as a dynamic guardian of the endothelial genome and facilitates rapid switching from quiescence to proliferation.

Original language	English
Journal	Cell Reports
DOIs	https://doi.org/10.1016/j.celrep.2022.111670
Publication status	Published - 15 Nov 2022

Keywords / Materials (for Non-textual outputs)

Long non-coding RNA
endothelial cells
replication protein A
quiescence
checkpoint control
Ataxia telangiectasia and Rad3-related

Access to Document

10.1016/j.celrep.2022.111670

CELL-REPORTS-D-22-01513_R5Accepted author manuscript, 25.3 MBLicence: Creative Commons: Attribution (CC-BY)

https://www.cell.com/cell-reports/fulltext/S2211-1247(22)01544-3?_returnURL=https%3A%2F%2Flinkinghub.elsevier.com%2Fretrieve%2Fpii%2FS2211124722015443%3Fshowall%3DtrueLicence: Creative Commons: Attribution (CC-BY)

Exploiting control of long non-coding RNAs to promote regenerative angiogenesis in the damaged myocardium
Baker, A., Brittan, M. & Rodor, J.
UK-based charities
1/01/22 → 31/12/25
Project: Research

Cite this

Oo, J. A., Pálfi, K., Warwick, T., Wittig, I., Prieto-Garcia, C., Matkovic, V., Tomašković, I., Boos, F., Izquierdo Ponce, J., Teichmann, T., Petriukov, K., Haydar, S., Maegdefessel, L., Wu, Z., Duc Pham, M., Krishnan, J., Baker, A. H., Günther, S., Ulrich, H. D., ... Brandes, R. P. (2022). Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2. Cell Reports. https://doi.org/10.1016/j.celrep.2022.111670

@article{7398e3acaf5c43b1bf14cfbea3a1ff1b,

title = "Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2",

abstract = "In healthy vessels, endothelial cells maintain a stable, differentiated and growth-arrested phenotype for years. Upon injury, a rapid phenotypic switch facilitates proliferation to restore tissue perfusion. Here we report the identification of the endothelial cell-enriched long non-coding RNA (lncRNA) PCAT19, which contributes to the proliferative switch and acts as a safeguard for the endothelialgenome. PCAT19 is enriched in confluent, quiescent endothelial cells and binds to the full replication protein A (RPA) complex in a DNA damage and cell cycle-related manner. Our results suggest that PCAT19 limits the phosphorylation of RPA2, primarily on the serine 33 (S33) residue, and thereby facilitates an appropriate DNA damage response while slowing cell cycle progression. Reduction in PCAT19 levels, either in response to loss of cell contacts or knockdown, promotes endothelial proliferation and angiogenesis. Collectively, PCAT19 acts as a dynamic guardian of the endothelial genome and facilitates rapid switching from quiescence to proliferation.",

keywords = "Long non-coding RNA, endothelial cells, replication protein A, quiescence, checkpoint control, Ataxia telangiectasia and Rad3-related",

author = "Oo, {James A.} and Katalin P{\'a}lfi and Timothy Warwick and Ilka Wittig and Cristian Prieto-Garcia and Vigor Matkovic and Ines Toma{\v s}kovi{\'c} and Frederike Boos and {Izquierdo Ponce}, Judit and Tom Teichmann and Kirill Petriukov and Shaza Haydar and Lars Maegdefessel and Zhiyuan Wu and {Duc Pham}, Minh and Jaya Krishnan and Baker, {Andrew H} and Stefan G{\"u}nther and Ulrich, {Helle D.} and Ivan Dikic and Leisegang, {Matthias S} and Brandes, {Ralf P}",

note = "Funding Information: We thank Jana Meisterknecht for help with mass spectrometry, Thomas B{\"o}ttger for help with the rISH-PLA method, Eva Wolf for help with RPA purification, and Nicolas Ja{\'e} for sharing RNA protein pull-down protocols. Support from the IMB Microscopy Core Facility is gratefully acknowledged. This work was supported by the Goethe University Frankfurt am Main, the DFG Excellence Cluster Cardiopulmonary Institute (CPI) EXS2026 , the Dr. Rolf M. Schwiete Stiftung, and the DFG Transregio TRR267 (Project-ID 403584255-TRR 267 ; TP A04 , TP A06 , TP B04 , TP B07 , and TP Z02 ). A.H.B. is supported by the British Heart Foundation Chair of Translational Cardiovascular Sciences. Work in I.D. lab is supported by the grants from Else Kroener Fresenius Stiftung and Dr. Rolf M. Schwiete Stiftung. R.P.B. and A.H.B. are supported by the BHF / DHF / DZHK grant “Exploiting endothelial long non-coding RNAs to promote regenerative angiogenesis in the damaged myocardium.” Publisher Copyright: {\textcopyright} 2022 The Author(s)",

year = "2022",

month = nov,

day = "15",

doi = "10.1016/j.celrep.2022.111670",

language = "English",

journal = "Cell Reports",

issn = "2211-1247",

publisher = "Cell Press",

}

Oo, JA, Pálfi, K, Warwick, T, Wittig, I, Prieto-Garcia, C, Matkovic, V, Tomašković, I, Boos, F, Izquierdo Ponce, J, Teichmann, T, Petriukov, K, Haydar, S, Maegdefessel, L, Wu, Z, Duc Pham, M, Krishnan, J, Baker, AH, Günther, S, Ulrich, HD, Dikic, I, Leisegang, MS & Brandes, RP 2022, 'Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2', Cell Reports. https://doi.org/10.1016/j.celrep.2022.111670

TY - JOUR

T1 - Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2

AU - Oo, James A.

AU - Pálfi, Katalin

AU - Warwick, Timothy

AU - Wittig, Ilka

AU - Prieto-Garcia, Cristian

AU - Matkovic, Vigor

AU - Tomašković, Ines

AU - Boos, Frederike

AU - Izquierdo Ponce, Judit

AU - Teichmann, Tom

AU - Petriukov, Kirill

AU - Haydar, Shaza

AU - Maegdefessel, Lars

AU - Wu, Zhiyuan

AU - Duc Pham, Minh

AU - Krishnan, Jaya

AU - Baker, Andrew H

AU - Günther, Stefan

AU - Ulrich, Helle D.

AU - Dikic, Ivan

AU - Leisegang, Matthias S

AU - Brandes, Ralf P

N1 - Funding Information: We thank Jana Meisterknecht for help with mass spectrometry, Thomas Böttger for help with the rISH-PLA method, Eva Wolf for help with RPA purification, and Nicolas Jaé for sharing RNA protein pull-down protocols. Support from the IMB Microscopy Core Facility is gratefully acknowledged. This work was supported by the Goethe University Frankfurt am Main, the DFG Excellence Cluster Cardiopulmonary Institute (CPI) EXS2026 , the Dr. Rolf M. Schwiete Stiftung, and the DFG Transregio TRR267 (Project-ID 403584255-TRR 267 ; TP A04 , TP A06 , TP B04 , TP B07 , and TP Z02 ). A.H.B. is supported by the British Heart Foundation Chair of Translational Cardiovascular Sciences. Work in I.D. lab is supported by the grants from Else Kroener Fresenius Stiftung and Dr. Rolf M. Schwiete Stiftung. R.P.B. and A.H.B. are supported by the BHF / DHF / DZHK grant “Exploiting endothelial long non-coding RNAs to promote regenerative angiogenesis in the damaged myocardium.” Publisher Copyright: © 2022 The Author(s)

PY - 2022/11/15

Y1 - 2022/11/15

N2 - In healthy vessels, endothelial cells maintain a stable, differentiated and growth-arrested phenotype for years. Upon injury, a rapid phenotypic switch facilitates proliferation to restore tissue perfusion. Here we report the identification of the endothelial cell-enriched long non-coding RNA (lncRNA) PCAT19, which contributes to the proliferative switch and acts as a safeguard for the endothelialgenome. PCAT19 is enriched in confluent, quiescent endothelial cells and binds to the full replication protein A (RPA) complex in a DNA damage and cell cycle-related manner. Our results suggest that PCAT19 limits the phosphorylation of RPA2, primarily on the serine 33 (S33) residue, and thereby facilitates an appropriate DNA damage response while slowing cell cycle progression. Reduction in PCAT19 levels, either in response to loss of cell contacts or knockdown, promotes endothelial proliferation and angiogenesis. Collectively, PCAT19 acts as a dynamic guardian of the endothelial genome and facilitates rapid switching from quiescence to proliferation.

AB - In healthy vessels, endothelial cells maintain a stable, differentiated and growth-arrested phenotype for years. Upon injury, a rapid phenotypic switch facilitates proliferation to restore tissue perfusion. Here we report the identification of the endothelial cell-enriched long non-coding RNA (lncRNA) PCAT19, which contributes to the proliferative switch and acts as a safeguard for the endothelialgenome. PCAT19 is enriched in confluent, quiescent endothelial cells and binds to the full replication protein A (RPA) complex in a DNA damage and cell cycle-related manner. Our results suggest that PCAT19 limits the phosphorylation of RPA2, primarily on the serine 33 (S33) residue, and thereby facilitates an appropriate DNA damage response while slowing cell cycle progression. Reduction in PCAT19 levels, either in response to loss of cell contacts or knockdown, promotes endothelial proliferation and angiogenesis. Collectively, PCAT19 acts as a dynamic guardian of the endothelial genome and facilitates rapid switching from quiescence to proliferation.

KW - Long non-coding RNA

KW - endothelial cells

KW - replication protein A

KW - quiescence

KW - checkpoint control

KW - Ataxia telangiectasia and Rad3-related

U2 - 10.1016/j.celrep.2022.111670

DO - 10.1016/j.celrep.2022.111670

M3 - Article

SN - 2211-1247

JO - Cell Reports

JF - Cell Reports

ER -

Long non-coding RNA PCAT19 safeguards DNA in quiescent endothelial cells by preventing uncontrolled phosphorylation of replication protein A2

Abstract / Description of output

Keywords / Materials (for Non-textual outputs)

Access to Document

Fingerprint

Projects

Exploiting control of long non-coding RNAs to promote regenerative angiogenesis in the damaged myocardium

Cite this