The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication

Min Jiang; Xie Xie; Xuefeng Zhu; Shan Jiang; Dusanka Milenkovic; Jelena Misic; Yonghong Shi; Nirwan Tandukar; Xinping Li; Ilian Atanassov; Louise Jenninger; Emily Hoberg; Sara Albarran-Gutierrez; Zsolt Szilagyi; Bertil Macao; Stefan J. Siira; Valerio Carelli; Jack D. Griffith; Claes M. Gustafsson; Thomas J. Nicholls; Aleksandra Filipovska; Nils Göran Larsson; Maria Falkenberg

doi:10.1126/sciadv.abf8631

The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication

Min Jiang, Xie Xie, Xuefeng Zhu, Shan Jiang, Dusanka Milenkovic, Jelena Misic, Yonghong Shi, Nirwan Tandukar, Xinping Li, Ilian Atanassov, Louise Jenninger, Emily Hoberg, Sara Albarran-Gutierrez, Zsolt Szilagyi, Bertil Macao, Stefan J. Siira, Valerio Carelli, Jack D. Griffith, Claes M. Gustafsson, Thomas J. NichollsAleksandra Filipovska, Nils Göran Larsson, Maria Falkenberg

UWA Centre for Medical Research (affiliated with the Harry Perkins Institute of Medical Research)

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8 Citations (Web of Science)

Abstract

We report a role for the mitochondrial single-stranded DNA binding protein (mtSSB) in regulating mitochondrial DNA (mtDNA) replication initiation in mammalian mitochondria. Transcription from the light-strand promoter (LSP) is required both for gene expression and for generating the RNA primers needed for initiation of mtDNA synthesis. In the absence of mtSSB, transcription from LSP is strongly up-regulated, but no replication primers are formed. Using deep sequencing in a mouse knockout model and biochemical reconstitution experiments with pure proteins, we find that mtSSB is necessary to restrict transcription initiation to optimize RNA primer formation at both origins of mtDNA replication. Last, we show that human pathological versions of mtSSB causing severe mitochondrial disease cannot efficiently support primer formation and initiation of mtDNA replication.

Original language	English
Article number	eabf8631
Journal	Science Advances
Volume	7
Issue number	27
DOIs	https://doi.org/10.1126/sciadv.abf8631
Publication status	Published - Jun 2021

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Jiang, M., Xie, X., Zhu, X., Jiang, S., Milenkovic, D., Misic, J., Shi, Y., Tandukar, N., Li, X., Atanassov, I., Jenninger, L., Hoberg, E., Albarran-Gutierrez, S., Szilagyi, Z., Macao, B., Siira, S. J., Carelli, V., Griffith, J. D., Gustafsson, C. M., ... Falkenberg, M. (2021). The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication. Science Advances, 7(27), [eabf8631]. https://doi.org/10.1126/sciadv.abf8631

Jiang, Min ; Xie, Xie ; Zhu, Xuefeng ; Jiang, Shan ; Milenkovic, Dusanka ; Misic, Jelena ; Shi, Yonghong ; Tandukar, Nirwan ; Li, Xinping ; Atanassov, Ilian ; Jenninger, Louise ; Hoberg, Emily ; Albarran-Gutierrez, Sara ; Szilagyi, Zsolt ; Macao, Bertil ; Siira, Stefan J. ; Carelli, Valerio ; Griffith, Jack D. ; Gustafsson, Claes M. ; Nicholls, Thomas J. ; Filipovska, Aleksandra ; Larsson, Nils Göran ; Falkenberg, Maria. / The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication. In: Science Advances. 2021 ; Vol. 7, No. 27.

@article{dc9a9b96cb704f44b4eb4ee10add05f4,

title = "The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication",

abstract = "We report a role for the mitochondrial single-stranded DNA binding protein (mtSSB) in regulating mitochondrial DNA (mtDNA) replication initiation in mammalian mitochondria. Transcription from the light-strand promoter (LSP) is required both for gene expression and for generating the RNA primers needed for initiation of mtDNA synthesis. In the absence of mtSSB, transcription from LSP is strongly up-regulated, but no replication primers are formed. Using deep sequencing in a mouse knockout model and biochemical reconstitution experiments with pure proteins, we find that mtSSB is necessary to restrict transcription initiation to optimize RNA primer formation at both origins of mtDNA replication. Last, we show that human pathological versions of mtSSB causing severe mitochondrial disease cannot efficiently support primer formation and initiation of mtDNA replication.",

author = "Min Jiang and Xie Xie and Xuefeng Zhu and Shan Jiang and Dusanka Milenkovic and Jelena Misic and Yonghong Shi and Nirwan Tandukar and Xinping Li and Ilian Atanassov and Louise Jenninger and Emily Hoberg and Sara Albarran-Gutierrez and Zsolt Szilagyi and Bertil Macao and Siira, {Stefan J.} and Valerio Carelli and Griffith, {Jack D.} and Gustafsson, {Claes M.} and Nicholls, {Thomas J.} and Aleksandra Filipovska and Larsson, {Nils G{\"o}ran} and Maria Falkenberg",

year = "2021",

month = jun,

doi = "10.1126/sciadv.abf8631",

language = "English",

volume = "7",

journal = "Science Advances",

issn = "2375-2548",

publisher = "AMER ASSOC ADVANCEMENT SCIENCE",

number = "27",

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Jiang, M, Xie, X, Zhu, X, Jiang, S, Milenkovic, D, Misic, J, Shi, Y, Tandukar, N, Li, X, Atanassov, I, Jenninger, L, Hoberg, E, Albarran-Gutierrez, S, Szilagyi, Z, Macao, B, Siira, SJ, Carelli, V, Griffith, JD, Gustafsson, CM, Nicholls, TJ, Filipovska, A, Larsson, NG & Falkenberg, M 2021, 'The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication', Science Advances, vol. 7, no. 27, eabf8631. https://doi.org/10.1126/sciadv.abf8631

The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication. / Jiang, Min; Xie, Xie; Zhu, Xuefeng; Jiang, Shan; Milenkovic, Dusanka; Misic, Jelena; Shi, Yonghong; Tandukar, Nirwan; Li, Xinping; Atanassov, Ilian; Jenninger, Louise; Hoberg, Emily; Albarran-Gutierrez, Sara; Szilagyi, Zsolt; Macao, Bertil; Siira, Stefan J.; Carelli, Valerio; Griffith, Jack D.; Gustafsson, Claes M.; Nicholls, Thomas J.; Filipovska, Aleksandra; Larsson, Nils Göran; Falkenberg, Maria.

In: Science Advances, Vol. 7, No. 27, eabf8631, 06.2021.

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

TY - JOUR

T1 - The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication

AU - Jiang, Min

AU - Xie, Xie

AU - Zhu, Xuefeng

AU - Jiang, Shan

AU - Milenkovic, Dusanka

AU - Misic, Jelena

AU - Shi, Yonghong

AU - Tandukar, Nirwan

AU - Li, Xinping

AU - Atanassov, Ilian

AU - Jenninger, Louise

AU - Hoberg, Emily

AU - Albarran-Gutierrez, Sara

AU - Szilagyi, Zsolt

AU - Macao, Bertil

AU - Siira, Stefan J.

AU - Carelli, Valerio

AU - Griffith, Jack D.

AU - Gustafsson, Claes M.

AU - Nicholls, Thomas J.

AU - Filipovska, Aleksandra

AU - Larsson, Nils Göran

AU - Falkenberg, Maria

PY - 2021/6

Y1 - 2021/6

N2 - We report a role for the mitochondrial single-stranded DNA binding protein (mtSSB) in regulating mitochondrial DNA (mtDNA) replication initiation in mammalian mitochondria. Transcription from the light-strand promoter (LSP) is required both for gene expression and for generating the RNA primers needed for initiation of mtDNA synthesis. In the absence of mtSSB, transcription from LSP is strongly up-regulated, but no replication primers are formed. Using deep sequencing in a mouse knockout model and biochemical reconstitution experiments with pure proteins, we find that mtSSB is necessary to restrict transcription initiation to optimize RNA primer formation at both origins of mtDNA replication. Last, we show that human pathological versions of mtSSB causing severe mitochondrial disease cannot efficiently support primer formation and initiation of mtDNA replication.

AB - We report a role for the mitochondrial single-stranded DNA binding protein (mtSSB) in regulating mitochondrial DNA (mtDNA) replication initiation in mammalian mitochondria. Transcription from the light-strand promoter (LSP) is required both for gene expression and for generating the RNA primers needed for initiation of mtDNA synthesis. In the absence of mtSSB, transcription from LSP is strongly up-regulated, but no replication primers are formed. Using deep sequencing in a mouse knockout model and biochemical reconstitution experiments with pure proteins, we find that mtSSB is necessary to restrict transcription initiation to optimize RNA primer formation at both origins of mtDNA replication. Last, we show that human pathological versions of mtSSB causing severe mitochondrial disease cannot efficiently support primer formation and initiation of mtDNA replication.

UR - http://www.scopus.com/inward/record.url?scp=85109333330&partnerID=8YFLogxK

U2 - 10.1126/sciadv.abf8631

DO - 10.1126/sciadv.abf8631

M3 - Article

C2 - 34215584

AN - SCOPUS:85109333330

VL - 7

JO - Science Advances

JF - Science Advances

SN - 2375-2548

IS - 27

M1 - eabf8631

ER -

The mitochondrial single-stranded DNA binding protein is essential for initiation of mtDNA replication

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