A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription
Extracellular signal-regulated kinase (ERK) regulates multiple cellular functions through distinct activation patterns. Genetically encoded fluorescent probes are instrumental in dissecting the ERK activity dynamics in living cells. Here we modified a previously reported Förster resonance energy tra...
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| Format: | Article |
| Language: | English |
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Japan Society for Cell Biology
2024-12-01
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| Series: | Cell Structure and Function |
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| Online Access: | https://www.jstage.jst.go.jp/article/csf/50/1/50_24070/_html/-char/en |
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| author | Zhang Weisheng Jun Nakayama Yukino Inomata Shigeki Higashiyama Toru Hiratsuka |
| author_facet | Zhang Weisheng Jun Nakayama Yukino Inomata Shigeki Higashiyama Toru Hiratsuka |
| author_sort | Zhang Weisheng |
| collection | DOAJ |
| description | Extracellular signal-regulated kinase (ERK) regulates multiple cellular functions through distinct activation patterns. Genetically encoded fluorescent probes are instrumental in dissecting the ERK activity dynamics in living cells. Here we modified a previously reported Förster resonance energy transfer (FRET) probe for ERK, EKAREN5 by replacing its mTurquoise2 and YPet sequences with mTurquoise-GL and a synonymous codon variant of YPet, respectively. The modified biosensor, EKAREN5-gl, showed an increased sensitivity to EGF-induced ERK activation responding to a very low dose (20 pg/ml) of EGF stimulation. We quantitatively characterized two FRET-based ERK probes, EKAREN5 and EKAREN5-gl, and a subcellular kinase translocation-based probe, ERK-KTR. We found the three biosensors differently respond to EGF stimulations with different intensity, duration, and latency. Furthermore, we investigated how the minimal EGF-induced ERK activation affects the downstream transcription in HeLa cells by comprehensive transcriptional analysis. We found the minimal ERK activation leads to a distinct transcriptional pattern from those induced by higher ERK activations. Our study highlights the significance of sensitive fluorescent probes to understand cellular signal dynamics and the role of minimal ERK activation in regulating transcription. Key words: fluorescent probe, ERK, FRET, KTR |
| format | Article |
| id | doaj-art-1207ef57e077402aba15c5809c5b9ac8 |
| institution | Kabale University |
| issn | 0386-7196 1347-3700 |
| language | English |
| publishDate | 2024-12-01 |
| publisher | Japan Society for Cell Biology |
| record_format | Article |
| series | Cell Structure and Function |
| spelling | doaj-art-1207ef57e077402aba15c5809c5b9ac82025-02-06T23:34:29ZengJapan Society for Cell BiologyCell Structure and Function0386-71961347-37002024-12-01501152410.1247/csf.24070csfA sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcriptionZhang Weisheng0Jun Nakayama1Yukino Inomata2Shigeki Higashiyama3Toru Hiratsuka4Department of Molecular Oncology, Graduate School of Medicine, Osaka UniversityDepartment of Oncogenesis and Growth Regulation, Research Center, Osaka International Cancer InstituteDepartment of Oncogenesis and Growth Regulation, Research Center, Osaka International Cancer InstituteDepartment of Oncogenesis and Growth Regulation, Research Center, Osaka International Cancer InstituteDepartment of Molecular Oncology, Graduate School of Medicine, Osaka UniversityExtracellular signal-regulated kinase (ERK) regulates multiple cellular functions through distinct activation patterns. Genetically encoded fluorescent probes are instrumental in dissecting the ERK activity dynamics in living cells. Here we modified a previously reported Förster resonance energy transfer (FRET) probe for ERK, EKAREN5 by replacing its mTurquoise2 and YPet sequences with mTurquoise-GL and a synonymous codon variant of YPet, respectively. The modified biosensor, EKAREN5-gl, showed an increased sensitivity to EGF-induced ERK activation responding to a very low dose (20 pg/ml) of EGF stimulation. We quantitatively characterized two FRET-based ERK probes, EKAREN5 and EKAREN5-gl, and a subcellular kinase translocation-based probe, ERK-KTR. We found the three biosensors differently respond to EGF stimulations with different intensity, duration, and latency. Furthermore, we investigated how the minimal EGF-induced ERK activation affects the downstream transcription in HeLa cells by comprehensive transcriptional analysis. We found the minimal ERK activation leads to a distinct transcriptional pattern from those induced by higher ERK activations. Our study highlights the significance of sensitive fluorescent probes to understand cellular signal dynamics and the role of minimal ERK activation in regulating transcription. Key words: fluorescent probe, ERK, FRET, KTRhttps://www.jstage.jst.go.jp/article/csf/50/1/50_24070/_html/-char/enfluorescent probeerkfretktr |
| spellingShingle | Zhang Weisheng Jun Nakayama Yukino Inomata Shigeki Higashiyama Toru Hiratsuka A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription Cell Structure and Function fluorescent probe erk fret ktr |
| title | A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription |
| title_full | A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription |
| title_fullStr | A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription |
| title_full_unstemmed | A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription |
| title_short | A sensitive ERK fluorescent probe reveals the significance of minimal EGF-induced transcription |
| title_sort | sensitive erk fluorescent probe reveals the significance of minimal egf induced transcription |
| topic | fluorescent probe erk fret ktr |
| url | https://www.jstage.jst.go.jp/article/csf/50/1/50_24070/_html/-char/en |
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