Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases
Plant-specific Rho-type GTPases (ROPs) are master regulators of cell polarity and development. Over the past 30 years, their localization and dynamics have been largely examined with fluorescent proteins fused at the amino terminus without investigating their impact on protein function. The moss Phy...
| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2024-12-01
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| Series: | Plant Signaling & Behavior |
| Subjects: | |
| Online Access: | http://dx.doi.org/10.1080/15592324.2024.2306790 |
| _version_ | 1797336033894858752 |
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| author | Jingtong Ruan Zihan Yin Peishan Yi |
| author_facet | Jingtong Ruan Zihan Yin Peishan Yi |
| author_sort | Jingtong Ruan |
| collection | DOAJ |
| description | Plant-specific Rho-type GTPases (ROPs) are master regulators of cell polarity and development. Over the past 30 years, their localization and dynamics have been largely examined with fluorescent proteins fused at the amino terminus without investigating their impact on protein function. The moss Physcomitrium patens genome encodes four rop genes. In this study, we introduce a fluorescent tag at the endogenous amino terminus of ROP4 in wild-type and rop1,2,3 triple mutant via homologous recombination and demonstrate that the fluorescent tag severely impairs ROP4 function and inhibits its localization on the plasma membrane. This phenotype is exacerbated in mutants lacking ROP-related GTPase-activating proteins. By comparing the localization of nonfunctional and functional ROP4 fusion reporters, we provide insight into the mechanism that governs the membrane association of ROPs. |
| first_indexed | 2024-03-08T08:48:17Z |
| format | Article |
| id | doaj.art-4ced33b9b40d49a7836c5f7e092dd7f4 |
| institution | Directory Open Access Journal |
| issn | 1559-2316 1559-2324 |
| language | English |
| last_indexed | 2024-03-08T08:48:17Z |
| publishDate | 2024-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | Plant Signaling & Behavior |
| spelling | doaj.art-4ced33b9b40d49a7836c5f7e092dd7f42024-02-01T14:39:33ZengTaylor & Francis GroupPlant Signaling & Behavior1559-23161559-23242024-12-0119110.1080/15592324.2024.23067902306790Effects of fluorescent tags and activity status on the membrane localization of ROP GTPasesJingtong Ruan0Zihan Yin1Peishan Yi2Sichuan UniversitySichuan UniversitySichuan UniversityPlant-specific Rho-type GTPases (ROPs) are master regulators of cell polarity and development. Over the past 30 years, their localization and dynamics have been largely examined with fluorescent proteins fused at the amino terminus without investigating their impact on protein function. The moss Physcomitrium patens genome encodes four rop genes. In this study, we introduce a fluorescent tag at the endogenous amino terminus of ROP4 in wild-type and rop1,2,3 triple mutant via homologous recombination and demonstrate that the fluorescent tag severely impairs ROP4 function and inhibits its localization on the plasma membrane. This phenotype is exacerbated in mutants lacking ROP-related GTPase-activating proteins. By comparing the localization of nonfunctional and functional ROP4 fusion reporters, we provide insight into the mechanism that governs the membrane association of ROPs.http://dx.doi.org/10.1080/15592324.2024.2306790cell polarityrop gtpasemossfluorescent protein taggingmembrane trafficking |
| spellingShingle | Jingtong Ruan Zihan Yin Peishan Yi Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases Plant Signaling & Behavior cell polarity rop gtpase moss fluorescent protein tagging membrane trafficking |
| title | Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases |
| title_full | Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases |
| title_fullStr | Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases |
| title_full_unstemmed | Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases |
| title_short | Effects of fluorescent tags and activity status on the membrane localization of ROP GTPases |
| title_sort | effects of fluorescent tags and activity status on the membrane localization of rop gtpases |
| topic | cell polarity rop gtpase moss fluorescent protein tagging membrane trafficking |
| url | http://dx.doi.org/10.1080/15592324.2024.2306790 |
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