The ancestral type of the R-RAS protein has oncogenic potential
Abstract Background The R-RAS2 is a small GTPase highly similar to classical RAS proteins at the regulatory and signaling levels. The high evolutionary conservation of R-RAS2, its links to basic cellular processes and its role in cancer, make R-RAS2 an interesting research topic. To elucidate the ev...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
BMC
2024-02-01
|
| Series: | Cellular & Molecular Biology Letters |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s11658-024-00546-0 |
| _version_ | 1797273922344845312 |
|---|---|
| author | Antea Talajić Kristina Dominko Marija Lončarić Andreja Ambriović-Ristov Helena Ćetković |
| author_facet | Antea Talajić Kristina Dominko Marija Lončarić Andreja Ambriović-Ristov Helena Ćetković |
| author_sort | Antea Talajić |
| collection | DOAJ |
| description | Abstract Background The R-RAS2 is a small GTPase highly similar to classical RAS proteins at the regulatory and signaling levels. The high evolutionary conservation of R-RAS2, its links to basic cellular processes and its role in cancer, make R-RAS2 an interesting research topic. To elucidate the evolutionary history of R-RAS proteins, we investigated and compared structural and functional properties of ancestral type R-RAS protein with human R-RAS2. Methods Bioinformatics analysis were used to elucidate the evolution of R-RAS proteins. Intrinsic GTPase activity of purified human and sponge proteins was analyzed with GTPase-GloTM Assay kit. The cell model consisted of human breast cancer cell lines MCF-7 and MDA-MB-231 transiently transfected with EsuRRAS2-like or HsaRRAS2. Biological characterization of R-RAS2 proteins was performed by Western blot on whole cell lysates or cell adhesion protein isolates, immunofluorescence and confocal microscopy, MTT test, colony formation assay, wound healing and Boyden chamber migration assays. Results We found that the single sponge R-RAS2-like gene/protein probably reflects the properties of the ancestral R-RAS protein that existed prior to duplications during the transition to Bilateria, and to Vertebrata. Biochemical characterization of sponge and human R-RAS2 showed that they have the same intrinsic GTPase activity and RNA binding properties. By testing cell proliferation, migration and colony forming efficiency in MDA-MB-231 human breast cancer cells, we showed that the ancestral type of the R-RAS protein, sponge R-RAS2-like, enhances their oncogenic potential, similar to human R-RAS2. In addition, sponge and human R-RAS2 were not found in focal adhesions, but both homologs play a role in their regulation by increasing talin1 and vinculin. Conclusions This study suggests that the ancestor of all animals possessed an R-RAS2-like protein with oncogenic properties similar to evolutionarily more recent versions of the protein, even before the appearance of true tissue and the origin of tumors. Therefore, we have unraveled the evolutionary history of R-RAS2 in metazoans and improved our knowledge of R-RAS2 properties, including its structure, regulation and function. |
| first_indexed | 2024-03-07T14:51:03Z |
| format | Article |
| id | doaj.art-eb610082c1004645855ce823dc4e737c |
| institution | Directory Open Access Journal |
| issn | 1689-1392 |
| language | English |
| last_indexed | 2024-03-07T14:51:03Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Cellular & Molecular Biology Letters |
| spelling | doaj.art-eb610082c1004645855ce823dc4e737c2024-03-05T19:44:22ZengBMCCellular & Molecular Biology Letters1689-13922024-02-0129112510.1186/s11658-024-00546-0The ancestral type of the R-RAS protein has oncogenic potentialAntea Talajić0Kristina Dominko1Marija Lončarić2Andreja Ambriović-Ristov3Helena Ćetković4Laboratory for Molecular Genetics, Division of Molecular Biology, Ruđer Bošković InstituteLaboratory for Molecular Genetics, Division of Molecular Biology, Ruđer Bošković InstituteLaboratory for Cell Biology and Signalling, Division of Molecular Biology, Ruđer Bošković InstituteLaboratory for Cell Biology and Signalling, Division of Molecular Biology, Ruđer Bošković InstituteLaboratory for Molecular Genetics, Division of Molecular Biology, Ruđer Bošković InstituteAbstract Background The R-RAS2 is a small GTPase highly similar to classical RAS proteins at the regulatory and signaling levels. The high evolutionary conservation of R-RAS2, its links to basic cellular processes and its role in cancer, make R-RAS2 an interesting research topic. To elucidate the evolutionary history of R-RAS proteins, we investigated and compared structural and functional properties of ancestral type R-RAS protein with human R-RAS2. Methods Bioinformatics analysis were used to elucidate the evolution of R-RAS proteins. Intrinsic GTPase activity of purified human and sponge proteins was analyzed with GTPase-GloTM Assay kit. The cell model consisted of human breast cancer cell lines MCF-7 and MDA-MB-231 transiently transfected with EsuRRAS2-like or HsaRRAS2. Biological characterization of R-RAS2 proteins was performed by Western blot on whole cell lysates or cell adhesion protein isolates, immunofluorescence and confocal microscopy, MTT test, colony formation assay, wound healing and Boyden chamber migration assays. Results We found that the single sponge R-RAS2-like gene/protein probably reflects the properties of the ancestral R-RAS protein that existed prior to duplications during the transition to Bilateria, and to Vertebrata. Biochemical characterization of sponge and human R-RAS2 showed that they have the same intrinsic GTPase activity and RNA binding properties. By testing cell proliferation, migration and colony forming efficiency in MDA-MB-231 human breast cancer cells, we showed that the ancestral type of the R-RAS protein, sponge R-RAS2-like, enhances their oncogenic potential, similar to human R-RAS2. In addition, sponge and human R-RAS2 were not found in focal adhesions, but both homologs play a role in their regulation by increasing talin1 and vinculin. Conclusions This study suggests that the ancestor of all animals possessed an R-RAS2-like protein with oncogenic properties similar to evolutionarily more recent versions of the protein, even before the appearance of true tissue and the origin of tumors. Therefore, we have unraveled the evolutionary history of R-RAS2 in metazoans and improved our knowledge of R-RAS2 properties, including its structure, regulation and function.https://doi.org/10.1186/s11658-024-00546-0CancerCell migrationCell proliferationEvolutionFocal adhesionIntracellular localization |
| spellingShingle | Antea Talajić Kristina Dominko Marija Lončarić Andreja Ambriović-Ristov Helena Ćetković The ancestral type of the R-RAS protein has oncogenic potential Cellular & Molecular Biology Letters Cancer Cell migration Cell proliferation Evolution Focal adhesion Intracellular localization |
| title | The ancestral type of the R-RAS protein has oncogenic potential |
| title_full | The ancestral type of the R-RAS protein has oncogenic potential |
| title_fullStr | The ancestral type of the R-RAS protein has oncogenic potential |
| title_full_unstemmed | The ancestral type of the R-RAS protein has oncogenic potential |
| title_short | The ancestral type of the R-RAS protein has oncogenic potential |
| title_sort | ancestral type of the r ras protein has oncogenic potential |
| topic | Cancer Cell migration Cell proliferation Evolution Focal adhesion Intracellular localization |
| url | https://doi.org/10.1186/s11658-024-00546-0 |
| work_keys_str_mv | AT anteatalajic theancestraltypeoftherrasproteinhasoncogenicpotential AT kristinadominko theancestraltypeoftherrasproteinhasoncogenicpotential AT marijaloncaric theancestraltypeoftherrasproteinhasoncogenicpotential AT andrejaambriovicristov theancestraltypeoftherrasproteinhasoncogenicpotential AT helenacetkovic theancestraltypeoftherrasproteinhasoncogenicpotential AT anteatalajic ancestraltypeoftherrasproteinhasoncogenicpotential AT kristinadominko ancestraltypeoftherrasproteinhasoncogenicpotential AT marijaloncaric ancestraltypeoftherrasproteinhasoncogenicpotential AT andrejaambriovicristov ancestraltypeoftherrasproteinhasoncogenicpotential AT helenacetkovic ancestraltypeoftherrasproteinhasoncogenicpotential |