Acetylation and phosphorylation of human TFAM regulate TFAM-DNA interactions via contrasting mechanisms

Maryam Hashemi Shabestari, Graeme A. King, Kees-Karel H. Taris, Ashutosh K. Pandey, Sundararajan Venkatesh, Jayapalraja Thilagavathi, Kamalendra Singh, Rama Krishna Koppisetti, Dmitry Temiakov, Wouter H. Roos, Carolyn K. Suzuki, Gijs J. L. Wuite

Research output: Contribution to JournalArticleAcademicpeer-review

Abstract

Mitochondrial transcription factor A (TFAM) is essential for the maintenance, expression and transmission of mitochondrial DNA (mtDNA). However, mechanisms for the post-translational regulation of TFAM are poorly understood. Here, we show that TFAM is lysine acetylated within its high-mobility-group box 1, a domain that can also be serine phosphorylated. Using bulk and single-molecule methods, we demonstrate that site-specific phosphoserine and acetyllysine mimics of human TFAM regulate its interaction with non-specific DNA through distinct kinetic pathways. We show that higher protein concentrations of both TFAM mimics are required to compact DNA to a similar extent as the wild-type. Compaction is thought to be crucial for regulating mtDNA segregation and expression. Moreover, we reveal that the reduced DNA binding affinity of the acetyl-lysine mimic arises from a lower on-rate, whereas the phosphoserine mimic displays both a decreased on-rate and an increased off-rate. Strikingly, the increased off-rate of the phosphoserine mimic is coupled to a significantly faster diffusion of TFAM on DNA. These findings indicate that acetylation and phosphorylation of TFAM can fine-tune TFAM-DNA binding affinity, to permit the discrete regulation of mtDNA dynamics. Furthermore, our results suggest that phosphorylation could additionally regulate transcription by altering the ability of TFAM to locate promoter sites.
Original languageEnglish
Pages (from-to)3633-3642
JournalNucleic Acids Research
Volume46
Issue number7
DOIs
Publication statusPublished - 20 Apr 2018

Cite this

Shabestari, Maryam Hashemi ; King, Graeme A. ; Taris, Kees-Karel H. ; Pandey, Ashutosh K. ; Venkatesh, Sundararajan ; Thilagavathi, Jayapalraja ; Singh, Kamalendra ; Koppisetti, Rama Krishna ; Temiakov, Dmitry ; Roos, Wouter H. ; Suzuki, Carolyn K. ; Wuite, Gijs J. L. / Acetylation and phosphorylation of human TFAM regulate TFAM-DNA interactions via contrasting mechanisms. In: Nucleic Acids Research. 2018 ; Vol. 46, No. 7. pp. 3633-3642.
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abstract = "Mitochondrial transcription factor A (TFAM) is essential for the maintenance, expression and transmission of mitochondrial DNA (mtDNA). However, mechanisms for the post-translational regulation of TFAM are poorly understood. Here, we show that TFAM is lysine acetylated within its high-mobility-group box 1, a domain that can also be serine phosphorylated. Using bulk and single-molecule methods, we demonstrate that site-specific phosphoserine and acetyllysine mimics of human TFAM regulate its interaction with non-specific DNA through distinct kinetic pathways. We show that higher protein concentrations of both TFAM mimics are required to compact DNA to a similar extent as the wild-type. Compaction is thought to be crucial for regulating mtDNA segregation and expression. Moreover, we reveal that the reduced DNA binding affinity of the acetyl-lysine mimic arises from a lower on-rate, whereas the phosphoserine mimic displays both a decreased on-rate and an increased off-rate. Strikingly, the increased off-rate of the phosphoserine mimic is coupled to a significantly faster diffusion of TFAM on DNA. These findings indicate that acetylation and phosphorylation of TFAM can fine-tune TFAM-DNA binding affinity, to permit the discrete regulation of mtDNA dynamics. Furthermore, our results suggest that phosphorylation could additionally regulate transcription by altering the ability of TFAM to locate promoter sites.",
author = "Shabestari, {Maryam Hashemi} and King, {Graeme A.} and Taris, {Kees-Karel H.} and Pandey, {Ashutosh K.} and Sundararajan Venkatesh and Jayapalraja Thilagavathi and Kamalendra Singh and Koppisetti, {Rama Krishna} and Dmitry Temiakov and Roos, {Wouter H.} and Suzuki, {Carolyn K.} and Wuite, {Gijs J. L.}",
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Acetylation and phosphorylation of human TFAM regulate TFAM-DNA interactions via contrasting mechanisms. / Shabestari, Maryam Hashemi; King, Graeme A.; Taris, Kees-Karel H.; Pandey, Ashutosh K.; Venkatesh, Sundararajan; Thilagavathi, Jayapalraja; Singh, Kamalendra; Koppisetti, Rama Krishna; Temiakov, Dmitry; Roos, Wouter H.; Suzuki, Carolyn K.; Wuite, Gijs J. L.

In: Nucleic Acids Research, Vol. 46, No. 7, 20.04.2018, p. 3633-3642.

Research output: Contribution to JournalArticleAcademicpeer-review

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T1 - Acetylation and phosphorylation of human TFAM regulate TFAM-DNA interactions via contrasting mechanisms

AU - Shabestari, Maryam Hashemi

AU - King, Graeme A.

AU - Taris, Kees-Karel H.

AU - Pandey, Ashutosh K.

AU - Venkatesh, Sundararajan

AU - Thilagavathi, Jayapalraja

AU - Singh, Kamalendra

AU - Koppisetti, Rama Krishna

AU - Temiakov, Dmitry

AU - Roos, Wouter H.

AU - Suzuki, Carolyn K.

AU - Wuite, Gijs J. L.

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N2 - Mitochondrial transcription factor A (TFAM) is essential for the maintenance, expression and transmission of mitochondrial DNA (mtDNA). However, mechanisms for the post-translational regulation of TFAM are poorly understood. Here, we show that TFAM is lysine acetylated within its high-mobility-group box 1, a domain that can also be serine phosphorylated. Using bulk and single-molecule methods, we demonstrate that site-specific phosphoserine and acetyllysine mimics of human TFAM regulate its interaction with non-specific DNA through distinct kinetic pathways. We show that higher protein concentrations of both TFAM mimics are required to compact DNA to a similar extent as the wild-type. Compaction is thought to be crucial for regulating mtDNA segregation and expression. Moreover, we reveal that the reduced DNA binding affinity of the acetyl-lysine mimic arises from a lower on-rate, whereas the phosphoserine mimic displays both a decreased on-rate and an increased off-rate. Strikingly, the increased off-rate of the phosphoserine mimic is coupled to a significantly faster diffusion of TFAM on DNA. These findings indicate that acetylation and phosphorylation of TFAM can fine-tune TFAM-DNA binding affinity, to permit the discrete regulation of mtDNA dynamics. Furthermore, our results suggest that phosphorylation could additionally regulate transcription by altering the ability of TFAM to locate promoter sites.

AB - Mitochondrial transcription factor A (TFAM) is essential for the maintenance, expression and transmission of mitochondrial DNA (mtDNA). However, mechanisms for the post-translational regulation of TFAM are poorly understood. Here, we show that TFAM is lysine acetylated within its high-mobility-group box 1, a domain that can also be serine phosphorylated. Using bulk and single-molecule methods, we demonstrate that site-specific phosphoserine and acetyllysine mimics of human TFAM regulate its interaction with non-specific DNA through distinct kinetic pathways. We show that higher protein concentrations of both TFAM mimics are required to compact DNA to a similar extent as the wild-type. Compaction is thought to be crucial for regulating mtDNA segregation and expression. Moreover, we reveal that the reduced DNA binding affinity of the acetyl-lysine mimic arises from a lower on-rate, whereas the phosphoserine mimic displays both a decreased on-rate and an increased off-rate. Strikingly, the increased off-rate of the phosphoserine mimic is coupled to a significantly faster diffusion of TFAM on DNA. These findings indicate that acetylation and phosphorylation of TFAM can fine-tune TFAM-DNA binding affinity, to permit the discrete regulation of mtDNA dynamics. Furthermore, our results suggest that phosphorylation could additionally regulate transcription by altering the ability of TFAM to locate promoter sites.

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DO - 10.1093/nar/gky204

M3 - Article

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EP - 3642

JO - Nucleic Acids Research

JF - Nucleic Acids Research

SN - 0305-1048

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ER -