Alternative translation initiation generates a functionally distinct isoform of the stress-activated kinase MK2

Philipp  Trulley; Goda Snieckute; Dorte B. Bekker-Jensen; Manoj B Menon; Robert  Freund; Alexey  Kotlyarov; Jesper Velgaard Olsen; Manuel D.  Diaz-Muñoz; Martin  Turner; Simon Holst Bekker-Jensen; Matthias Gaestel; Christopher Tiedje

doi:10.1101/429696

Alternative translation initiation generates a functionally distinct isoform of the stress-activated kinase MK2

Philipp Trulley, Goda Snieckute, Dorte B. Bekker-Jensen, Manoj B Menon, Robert Freund, Alexey Kotlyarov, Jesper Velgaard Olsen, Manuel D. Diaz-Muñoz, Martin Turner, Simon Holst Bekker-Jensen, Matthias Gaestel, Christopher Tiedje

Abstract

Shaping of the proteome by alternative translation is an important mechanism of post-transcriptional gene regulation. It can lead to the expression of multiple protein isoforms originating from the same mRNA. Here we show that a novel, abundant and long isoform of the stress/p38MAPK-activated kinase MK2, a key regulator of transcription, migration, death signaling and post-transcriptional gene regulation, is constitutively translated from an alternative CUG translation initiation start site located in the 5′UTR of its mRNA. GC-rich sequences and putative G-quadruplex structures influence the usage of that codon as a translation initiation start site and the RNA helicase eIF4A1 is needed to ensure alternative isoform translation. We recapitulated the usage of the alternative start codon and determined the molecular properties of the short and a long MK2 isoforms. Phenotypically, only the short isoform phosphorylated Hsp27, supported migration and stress-induced immediate early gene (IEG) expression. Interaction profiling by quantitative mass-spectrometry revealed short isoform-specific binding partners that were associated with migration. In contrast, the long isoform contains additional putative phosphorylation sites in its unique N-terminus. In sum, our data reveal a longer and previously non-described isoform of MK2 with distinct physiological properties originating from alternative translation.

Original language	English
Journal	bioRxiv
Number of pages	46
DOIs	https://doi.org/10.1101/429696
Publication status	Published - 2018

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@article{5afc7589fcb144fb9fe759a227add18a,

title = "Alternative translation initiation generates a functionally distinct isoform of the stress-activated kinase MK2",

abstract = "Shaping of the proteome by alternative translation is an important mechanism of post-transcriptional gene regulation. It can lead to the expression of multiple protein isoforms originating from the same mRNA. Here we show that a novel, abundant and long isoform of the stress/p38MAPK-activated kinase MK2, a key regulator of transcription, migration, death signaling and post-transcriptional gene regulation, is constitutively translated from an alternative CUG translation initiation start site located in the 5′UTR of its mRNA. GC-rich sequences and putative G-quadruplex structures influence the usage of that codon as a translation initiation start site and the RNA helicase eIF4A1 is needed to ensure alternative isoform translation. We recapitulated the usage of the alternative start codon and determined the molecular properties of the short and a long MK2 isoforms. Phenotypically, only the short isoform phosphorylated Hsp27, supported migration and stress-induced immediate early gene (IEG) expression. Interaction profiling by quantitative mass-spectrometry revealed short isoform-specific binding partners that were associated with migration. In contrast, the long isoform contains additional putative phosphorylation sites in its unique N-terminus. In sum, our data reveal a longer and previously non-described isoform of MK2 with distinct physiological properties originating from alternative translation.",

author = "Philipp Trulley and Goda Snieckute and Bekker-Jensen, {Dorte B.} and Menon, {Manoj B} and Robert Freund and Alexey Kotlyarov and Olsen, {Jesper Velgaard} and Diaz-Mu{\~n}oz, {Manuel D.} and Martin Turner and Bekker-Jensen, {Simon Holst} and Matthias Gaestel and Christopher Tiedje",

year = "2018",

doi = "10.1101/429696",

language = "English",

journal = "bioRxiv",

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TY - JOUR

T1 - Alternative translation initiation generates a functionally distinct isoform of the stress-activated kinase MK2

AU - Trulley, Philipp

AU - Snieckute, Goda

AU - Bekker-Jensen, Dorte B.

AU - Menon, Manoj B

AU - Freund, Robert

AU - Kotlyarov, Alexey

AU - Olsen, Jesper Velgaard

AU - Diaz-Muñoz, Manuel D.

AU - Turner, Martin

AU - Bekker-Jensen, Simon Holst

AU - Gaestel, Matthias

AU - Tiedje, Christopher

PY - 2018

Y1 - 2018

N2 - Shaping of the proteome by alternative translation is an important mechanism of post-transcriptional gene regulation. It can lead to the expression of multiple protein isoforms originating from the same mRNA. Here we show that a novel, abundant and long isoform of the stress/p38MAPK-activated kinase MK2, a key regulator of transcription, migration, death signaling and post-transcriptional gene regulation, is constitutively translated from an alternative CUG translation initiation start site located in the 5′UTR of its mRNA. GC-rich sequences and putative G-quadruplex structures influence the usage of that codon as a translation initiation start site and the RNA helicase eIF4A1 is needed to ensure alternative isoform translation. We recapitulated the usage of the alternative start codon and determined the molecular properties of the short and a long MK2 isoforms. Phenotypically, only the short isoform phosphorylated Hsp27, supported migration and stress-induced immediate early gene (IEG) expression. Interaction profiling by quantitative mass-spectrometry revealed short isoform-specific binding partners that were associated with migration. In contrast, the long isoform contains additional putative phosphorylation sites in its unique N-terminus. In sum, our data reveal a longer and previously non-described isoform of MK2 with distinct physiological properties originating from alternative translation.

AB - Shaping of the proteome by alternative translation is an important mechanism of post-transcriptional gene regulation. It can lead to the expression of multiple protein isoforms originating from the same mRNA. Here we show that a novel, abundant and long isoform of the stress/p38MAPK-activated kinase MK2, a key regulator of transcription, migration, death signaling and post-transcriptional gene regulation, is constitutively translated from an alternative CUG translation initiation start site located in the 5′UTR of its mRNA. GC-rich sequences and putative G-quadruplex structures influence the usage of that codon as a translation initiation start site and the RNA helicase eIF4A1 is needed to ensure alternative isoform translation. We recapitulated the usage of the alternative start codon and determined the molecular properties of the short and a long MK2 isoforms. Phenotypically, only the short isoform phosphorylated Hsp27, supported migration and stress-induced immediate early gene (IEG) expression. Interaction profiling by quantitative mass-spectrometry revealed short isoform-specific binding partners that were associated with migration. In contrast, the long isoform contains additional putative phosphorylation sites in its unique N-terminus. In sum, our data reveal a longer and previously non-described isoform of MK2 with distinct physiological properties originating from alternative translation.

U2 - 10.1101/429696

DO - 10.1101/429696

M3 - Journal article

JO - bioRxiv

JF - bioRxiv

ER -

Alternative translation initiation generates a functionally distinct isoform of the stress-activated kinase MK2

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