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JK"],"technology_type":["Proteogenomics","Mass Spectrometry"],"software":[""],"submitter_keywords":["Chlamydomonas reinhardtii"],"full_dataset_link":["https://www.ebi.ac.uk/pride/archive/projects/PXD079084"],"tissue":["Cell Culture"],"sample_protocol":["For the in vitro kinase assay, the protein mixtures were incubated at 30 °C for 30 min in kinase buffer (20 mM Tris HCl, pH 7.5, 0.5 mM CaCl2, 2.5 mM MgCl2, 2.5 mM MnCl2, 1 mM DTT, 0.1 mM ATP). Reactions were terminated by the addition of 5× SDS loading buffer. Proteins were separated by SDS-PAGE, with total protein loading visualized by CBB staining."],"repository":["Pride"],"quantification_method":["Not available"],"modification":[""],"data_protocol":["The gel bands of interest were excised and subjected to in-gel digestion. Proteins were reduced and alkylated in 50 mM ammonium bicarbonate and then digested overnight at 37°C with trypsin at an enzyme-to-substrate ratio of 1:50. The resulting peptides were extracted twice with 1% formic acid in 50% aqueous acetonitrile and dried under vacuum. For LC-MS/MS analysis, the peptides were reconstituted in 0.1% formic acid and loaded automatically using a Bruker nanoElute UHPLC system onto a 25 cm analytical column (Aurora series, CSI; 75 µm inner diameter, 1.6 µm C18, IonOpticks) equipped with an integrated emitter. Peptides were separated over a 60-minute gradient at a constant flow rate of 300 nL/min using mobile phase A (water with 0.1% formic acid) and mobile phase B (acetonitrile with 0.1% formic acid). The gradient started at 2% B, increased to 22% B over 45 minutes, rose to 35% B by 50 minutes, and was held at 80% B from 55 to 60 minutes. Eluted peptides were ionized using a CaptiveSpray nano-electrospray source (Bruker Daltonics) and analyzed on a timsTOF Pro 2 instrument (Bruker Daltonics) operated in data-dependent acquisition parallel accumulation–serial fragmentation (dda-PASEF) mode. The acquisition method covered an m/z range of 300–1500 and an ion mobility (1/K₀) range of 0.75–1.3, with a ramp time of 166 ms. Raw data files were processed using Bruker Data Analysis 4.3 and Proteome Discoverer 2.5. Database searching was performed against the Chlamydomonas reinhardtii proteome (downloaded from UniProtKB) using the Sequest search algorithm. Search parameters included a precursor mass tolerance of 10 ppm, a fragment ion tolerance of 0.02 Da, and allowance for up to two missed cleavages. Carbamidomethylation of cysteine was set as a fixed modification, while oxidation of methionine and phosphorylation of serine, threonine, and tyrosine were included as variable modifications. Peptide identifications were filtered to achieve a 1% false discovery rate (FDR)."],"omics_type":["Proteomics"],"labhead":["xiaobo li"],"instrument_platform":[""],"labhead_affiliation":["Professor"],"submission_type":["PARTIAL"],"species":["Chlamydomonas Reinhardtii"],"publication":["10.1111/NPH.71438"],"submitter_mail":["guokangning@westlake.edu.cn"],"submitter_affiliation":["Westlake University"],"submitter_country":["China"],"additional_accession":[]},"is_claimable":false,"name":"A RAF–SnRK2–PP2C functional module regulates cell survival in the chlorophyte Chlamydomonas reinhardtii","description":"The SNF1-related protein kinase 2 (SnRK2) pathway is a central regulator of abiotic stress signaling in land plants; however, its evolutionary origins and functional conservation across the green lineage remain poorly understood. Land plants evolved from streptophyte algae, whereas the chlorophyte algae, including the widely studied model Chlamydomonas reinhardtii, represent an older lineage that diverged from streptophytes over one billion years ago. Here, we identify and characterize a functional SnRK2 signaling module in Chlamydomonas reinhardtii. We show that the isoform SnRK2.7 is required for osmotic stress tolerance and general cellular viability, thus offering an alternative model for dissecting SnRK2 functions in algae. SnRK2.7 localizes to the contractile vacuole, an osmoregulatory organelle lost during streptophyte evolution, revealing a lineage-specific functional adaptation. Together with MAPKKK3, a B1/B3-RAF kinase, and the clade A protein phosphatase PP2C3, these components constitute a SnRK2 signaling module in Chlamydomonas reinhardtii. Our findings demonstrate the presence of a functional SnRK2 pathway in a chlorophyte alga, indicating that core components were established early in the green lineage. This work provides a foundation for comparative studies across green plants and underscores the need for broader taxonomic sampling to reconstruct the ancestral signaling networks underlying stress adaptation.","dates":{"publication":"2026-07-02","submission":"2026-05-31"},"accession":"PXD079084","cross_references":{"TAXONOMY":["NEWT:330879","NEWT:377960","NEWT:2042546","NEWT:259447","NEWT:295546","NCBITaxon:2719036","NCBITaxon:1280","NEWT:112503","NEWT:1129","NEWT:309807","NEWT:59753","NEWT:89184","NEWT:309800","NEWT:281395","NEWT:1211601","NEWT:876138","NEWT:44271","NEWT:193516","NEWT:1073882","NEWT:475174","NEWT:111205","NEWT:1117","NEWT:498257","NEWT:10036","NEWT:1590","NEWT:10034","NEWT:661410","NEWT:638632","NEWT:224326","NEWT:376619","NCBITaxon:79857","NEWT:1096976","NEWT:1589","NEWT:135622","NEWT:35786","NEWT:1580","NEWT:399784","NEWT:96731","NEWT:656064","NEWT:383379","NEWT:418106","NEWT:10029","NEWT:913645","NEWT:641809","NEWT:38815","NEWT:317447","NEWT:4688","NEWT:111225","NEWT:7719","NEWT:868565","NEWT:135674","NEWT:79329","NEWT:30069","NEWT:12637","NEWT:59729","NEWT:2164133","NEWT:295105","NEWT:108061","NEWT:60711","NEWT:224308","NEWT:3347","NEWT:160621","NEWT:212790","NEWT:110368","NEWT:1310161","NEWT:77133","NEWT:145481","NEWT:1310165","NEWT:29058","NCBITaxon:79824","NEWT:1912919","NEWT:44688","NEWT:44689","NEWT:498211","NEWT:347256","NEWT:5518","NEWT:398007","NEWT:1527468","NEWT:498217","NEWT:498216","NEWT:11320","NEWT:246196","NEWT:246197","NEWT:145458","NEWT:44685","NEWT:161934","NEWT:1148","NEWT:5508","NEWT:3329","NEWT:419481","NEWT:5507","NEWT:410661","NEWT:1140","NCBITaxon:2157","NEWT:1143","NEWT:1287689","NEWT:1094343","NEWT:1462472","NEWT:1336795","NEWT:644042","NEWT:1182590","NEWT:3712","NEWT:3711","NEWT:270643","NEWT:2065263","NEWT:177437","NEWT:10418","NEWT:118698","NEWT:1616117","NEWT:118696","NEWT:34865","NEWT:52283","NEWT:284812","NEWT:8175","NEWT:43330","NEWT:1603293","NEWT:44664","NEWT:43346","NEWT:3702","NEWT:1245466","NEWT:244366","NEWT:1246791","NEWT:118694","NEWT:2850","NEWT:118691","NEWT:34871","NEWT:33548","NEWT:52299","NEWT:8165","NEWT:96794","NEWT:3708","NEWT:332648","NEWT:44670","NEWT:536231","NEWT:376219","NEWT:219813","NEWT:1510","NEWT:460519","NEWT:1515","NEWT:572307","NEWT:1432138","NEWT:1424507","NEWT:1194599","NEWT:272844","NEWT:1303443","NEWT:9483","NEWT:485","NEWT:56636","NEWT:2709072","NEWT:2853422","NEWT:1679718","NEWT:480","NEWT:67767","NEWT:46835","NEWT:109757","NEWT:582580","NEWT:1185650","NEWT:216778","NEWT:294607","NEWT:1502","NEWT:128017","NEWT:376686","NEWT:95486","NEWT:1883446","NEWT:1233435","NEWT:109760","NEWT:29031","NEWT:317047","NEWT:460523","NEWT:235443","NEWT:1757312","NEWT:85694","NEWT:108458","NEWT:5936","NEWT:320637","NEWT:1078283","NEWT:3750","NEWT:983964","NEWT:11706","NEWT:32644","NEWT:527796","NEWT:499175","NEWT:109779","NEWT:3745","NEWT:1715989","NCBITaxon:4751","NEWT:3747","NEWT:1116234","NEWT:1255228","NEWT:410289","NEWT:373153","NEWT:472","NEWT:940825","NEWT:529507","NEWT:1071661","NEWT:470","NEWT:5911","NCBITaxon:50557","NEWT:39251","NEWT:29491","NEWT:101841","NEWT:446","NEWT:153481","NEWT:2014887","NEWT:33952","NEWT:445","NEWT:153009","NEWT:261756","NEWT:166313","NEWT:63366","NEWT:63367","NEWT:215402","NEWT:1547","NEWT:906914","NEWT:9031","NEWT:27292","NEWT:1050722","NEWT:108931","NEWT:1293497","NEWT:1055524","NEWT:150475","NEWT:267872","NEWT:381666","NEWT:172269","NEWT:9534","NEWT:5180","NEWT:256737","NEWT:9541","NEWT:8694","NEWT:33936","NEWT:8692","NEWT:2903","NEWT:185579","NEWT:13076","NEWT:1006581","NEWT:8673","NEWT:33940","NEWT:550","NEWT:554","NEWT:451516","NEWT:552","NEWT:1325291","NEWT:36185","NEWT:1054211","NEWT:1225786","NEWT:51338","NEWT:575412","NEWT:28112","NEWT:6493","NEWT:6494","NEWT:6491","NEWT:507601","NEWT:520","NEWT:186441","NEWT:643680","NEWT:214092","NCBITaxon:6157","NEWT:13095","NEWT:162425","NEWT:104105","NEWT:216257","NEWT:9986","NEWT:8654","NEWT:8658","NEWT:1268063","NEWT:8657","NEWT:9983","NEWT:8655","NEWT:5147","NEWT:28108","NEWT:28104","NEWT:407821","NEWT:568703","NCBITaxon:2","NEWT:568708","NEWT:986","NEWT:52641","NEWT:28532","NEWT:353152","NEWT:298176","NEWT:227377","NEWT:40674","NEWT:1194669","NEWT:51329","NEWT:443906","NEWT:519","NEWT:2510939","NEWT:7394","NEWT:6063","NEWT:1328388","NEWT:1548728","NEWT:667127","NEWT:9557","NEWT:377586","NEWT:300641","NEWT:39655","NEWT:9554","NEWT:38323","NEWT:475932","NEWT:256318","NEWT:206411","NCBITaxon:6191","NEWT:229533","NEWT:214053","NEWT:80863","NEWT:90675","NEWT:52638","NEWT:57075","NEWT:1666905","NEWT:8697","NEWT:8695","NEWT:884204","NEWT:1123869","NEWT:9544","NEWT:9545","NEWT:979","NEWT:7370","NEWT:83906","NEWT:99488","NEWT:1134506","NEWT:255470","NEWT:38783","NEWT:6426","NEWT:33090","NEWT:9935","NEWT:287889","NEWT:305959","NEWT:13489","NEWT:29829","NEWT:92867","NEWT:92866","NCBITaxon:3055","NEWT:51750","NEWT:202950","NEWT:295027","NCBITaxon:11320","NEWT:632957","NEWT:9925","NCBITaxon:9606","NEWT:90690","NEWT:1436183","NEWT:300273","NEWT:4232","NEWT:416348","NEWT:11298","NEWT:4236","NEWT:7742","NEWT:196627","NEWT:200308","NEWT:200302","NEWT:870435","NEWT:9913","NEWT:9915","NEWT:105841","NEWT:2666255","NEWT:999810","NCBITaxon:5693","NEWT:28995","NEWT: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