{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,18]],"date-time":"2026-01-18T05:07:40Z","timestamp":1768712860763,"version":"3.49.0"},"reference-count":44,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,4,29]],"date-time":"2021-04-29T00:00:00Z","timestamp":1619654400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Chinese National Key R &amp; D Project for Synthetic Biology","award":["2018YFA0902500"],"award-info":[{"award-number":["2018YFA0902500"]}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["41876188, 31700309"],"award-info":[{"award-number":["41876188, 31700309"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Shenzhen Basic Research Projects","award":["JCYJ20180507182405562"],"award-info":[{"award-number":["JCYJ20180507182405562"]}]},{"name":"Guangxi Innovation Drive Development Special Fund","award":["Gui Ke AA18242047"],"award-info":[{"award-number":["Gui Ke AA18242047"]}]},{"name":"Funding for the Construction of Southern Marine Science and Engineering Guangdong Labora-tory","award":["Guangzhou"],"award-info":[{"award-number":["Guangzhou"]}]},{"name":"Grant Plan for Demonstration City Project for Marine Economic Development in Shenzhen to Dr. Zhangli HU.","award":["No.86"],"award-info":[{"award-number":["No.86"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Microalgae-based biodiesel production has many advantages over crude oil extraction and refinement, thus attracting more and more concern. Protein ubiquitination is a crucial mechanism in eukaryotes to regulate physiological responses and cell development, which is highly related to algal biodiesel production. Cullins as the molecular base of cullin-RING E3 ubiquitin ligases (CRLs), which are the largest known class of ubiquitin ligases, control the life activities of eukaryotic cells. Here, three cullins (CrCULs) in the green microalgae Chlamydomonas reinhardtii were identified and characterized. To investigate the roles of CrCULs in lipid metabolism, the gene expression profiles of CrCULs under nutrition starvation were examined. Except for down-regulation under nitrogen starvation, the CrCUL3 gene was induced by sulfur and iron starvation. CrCUL2 seemed insensitive to nitrogen and sulfur starvation because it only had changes after treatment for eight days. CrCUL4 exhibited an expression peak after nitrogen starvation for two days but this declined with time. All CrCULs expressions significantly increased under iron deficiency at two and four days but decreased thereafter. The silencing of CrCUL2 and CrCUL4 expression using RNAi (RNA interference) resulted in biomass decline and lipids increase but an increase of 20% and 28% in lipid content after growth for 10 days, respectively. In CrCUL2 and CrCUL4 RNAi lines, the content of fatty acids, especially C16:0 and C18:0, notably increased as well. However, the lipid content and fatty acids of the CrCUL3 RNAi strain slightly changed. Moreover, the subcellular localization of CrCUL4 showed a nuclear distribution pattern. These results suggest CrCUL2 and CrCUL4 are regulators for lipid accumulation in C. reinhardtii. This study may offer an important complement of lipid biosynthesis in microalgae.<\/jats:p>","DOI":"10.3390\/ijms22094695","type":"journal-article","created":{"date-parts":[[2021,4,29]],"date-time":"2021-04-29T04:30:11Z","timestamp":1619670611000},"page":"4695","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["The Roles of Cullins E3 Ubiquitin Ligases in the Lipid Biosynthesis of the Green Microalgae Chlamydomonas reinhardtii"],"prefix":"10.3390","volume":"22","author":[{"given":"Qiulan","family":"Luo","sequence":"first","affiliation":[{"name":"School of Life Sciences and Food Engineering, Hanshan Normal University, Chaozhou 521041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xianghui","family":"Zou","sequence":"additional","affiliation":[{"name":"School of Life Sciences and Food Engineering, Hanshan Normal University, Chaozhou 521041, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Chaogang","family":"Wang","sequence":"additional","affiliation":[{"name":"Guangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen Engineering Laboratory for Marine Algal Biotechnology, Longhua Innovation Institute for Biotechnology, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yajun","family":"Li","sequence":"additional","affiliation":[{"name":"Hainan Provincial Key Laboratory for Functional Components Research and Utilization of Marine Bio-Resources, Institute of Tropical Bioscience and Biotechnology, Hainan Academy of Tropical Agricultural Resource, Chinese Academy of Tropical Agricultural Sciences, Haikou 517010, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2596-8515","authenticated-orcid":false,"given":"Zhangli","family":"Hu","sequence":"additional","affiliation":[{"name":"Guangdong Technology Research Center for Marine Algal Bioengineering, Guangdong Provincial Key Laboratory for Plant Epigenetics, Shenzhen Engineering Laboratory for Marine Algal Biotechnology, Longhua Innovation Institute for Biotechnology, College of Life Sciences and Oceanography, Shenzhen University, Shenzhen 518060, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,29]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.rser.2018.05.012","article-title":"Production of biofuels from microalgae\u2014A review on cultivation, harvesting, lipid extraction, and numerous applications of microalgae","volume":"94","author":"Enamala","year":"2018","journal-title":"Renew. Sustain. Energy Rev."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"343","DOI":"10.4161\/bioe.21427","article-title":"Nitrogen and sulfur deprivation differentiate lipid accumulation targets of Chlamydomonas reinhardtii","volume":"3","author":"Cakmak","year":"2012","journal-title":"Bioengineered"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3708","DOI":"10.1105\/tpc.112.100701","article-title":"Phospholipid:diacylglycerol acyltransferase is a multifunctional enzyme involved in membrane lipid turnover and degradation while synthesizing triacylglycerol in the unicellular green microalga Chlamydomonas reinhardtii","volume":"24","author":"Yoon","year":"2012","journal-title":"Plant Cell"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2083","DOI":"10.1104\/pp.17.00491","article-title":"Two glycerol-3-phosphate dehydrogenases from Chlamydomonas have distinct roles in lipid metabolism","volume":"174","author":"Driver","year":"2017","journal-title":"Plant Physiol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2364","DOI":"10.1016\/j.febslet.2013.06.002","article-title":"Functional study of diacylglycerol acyltransferase type 2 family in Chlamydomonas reinhardtii","volume":"587","author":"Hung","year":"2013","journal-title":"FEBS Lett."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1007\/s00284-012-0095-6","article-title":"In silico cloning and characterization of the glycerol-3-phosphate dehydrogenase (GPDH) gene family in the green microalga Chlamydomonas reinhardtii","volume":"64","year":"2012","journal-title":"Curr. Microbiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1093\/mp\/sss040","article-title":"The roles of acyl-CoA: Diacylglycerol acyltransferase 2 genes in the biosynthesis of triacylglycerols by the green algae Chlamydomonas reinhardtii","volume":"5","author":"Deng","year":"2012","journal-title":"Mol. Plant."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1016\/j.jbiotec.2012.04.006","article-title":"Functional analysis of three type-2 DGAT homologue genes for triacylglycerol production in the green microalga Chlamydomonas reinhardtii","volume":"162","author":"Russa","year":"2012","journal-title":"J. Biotechnol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"60","DOI":"10.1016\/j.jbiotec.2017.03.004","article-title":"Expression of the heterologous Dunaliella tertiolecta fatty acyl-ACP thioesterase leads to increased lipid production in Chlamydomonas reinhardtii","volume":"247","author":"Tan","year":"2017","journal-title":"J. Biotechnol."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1527","DOI":"10.1016\/j.biortech.2017.04.111","article-title":"CRISPRi mediated phosphoenolpyruvate carboxylase regulation to enhance the production of lipid in Chlamydomonas reinhardtii","volume":"245","author":"Kao","year":"2017","journal-title":"Bioresour. Technol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1007\/s10529-019-02715-0","article-title":"Overexpression of acetyl-CoA carboxylase increases fatty acid production in the green alga Chlamydomonas reinhardtii","volume":"41","author":"Chen","year":"2019","journal-title":"Biotechnol. Lett."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1737","DOI":"10.1104\/pp.110.165159","article-title":"Changes in transcript abundance in Chlamydomonas reinhardtii following nitrogen deprivation predict diversion of metabolism","volume":"154","author":"Miller","year":"2010","journal-title":"Plant Physiol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"370","DOI":"10.1016\/j.febslet.2012.12.020","article-title":"Lipid droplet synthesis is limited by acetate availability in starchless mutant of Chlamydomonas reinhardtii","volume":"587","author":"Ramanan","year":"2013","journal-title":"FEBS Lett."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1146\/annurev.arplant.55.031903.141801","article-title":"The ubiquitin 26S proteasome proteolytic pathway","volume":"55","author":"Smalle","year":"2004","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"1763","DOI":"10.1126\/science.1123374","article-title":"TRB3 links the E3 ubiquitin ligase COP1 to lipid metabolism","volume":"312","author":"Qi","year":"2006","journal-title":"Science"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.ceb.2008.05.009","article-title":"Cell biology of lipid droplets","volume":"20","author":"Thiele","year":"2008","journal-title":"Curr. Opin. Cell. Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1146\/annurev-arplant-042809-112256","article-title":"The Cullin-RING ubiquitin-protein ligases","volume":"62","author":"Vierstra","year":"2011","journal-title":"Annu. Rev. Plant Biol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Mar\u00edn, I. (2009). Diversification of the cullin family. BMC Evol. Biol., 9.","DOI":"10.1186\/1471-2148-9-267"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"15275","DOI":"10.1073\/pnas.1006941107","article-title":"Cullin 4-ring finger-ligase plays a key role in the control of endoreplication cycles in Arabidopsis trichomes","volume":"107","author":"Roodbarkelari","year":"2010","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"16601","DOI":"10.1073\/pnas.252646399","article-title":"CUL7: A DOC domain-containing cullin selectively binds Skp1.Fbx29 to form an SCF-like complex","volume":"99","author":"Dias","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Yin, C.Y., Thilo, H., and Karl-Wilhelm, K. (2012). Mechanism of Cullin3 E3 Ubiquitin Ligase Dimerization. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0041350"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"731","DOI":"10.1038\/emboj.2010.359","article-title":"The Arabidopsis CUL4-DDB1 complex interacts with MSI1 and is required to maintain MEDEA parental imprinting","volume":"30","author":"Dumbliauskas","year":"2011","journal-title":"EMBO J."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"34","DOI":"10.1038\/s41477-018-0332-5","article-title":"Algal photoprotection is regulated by the E3 ligase CUL4-DDB1DET1","volume":"5","author":"Aihara","year":"2018","journal-title":"Nat. Plants"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"220","DOI":"10.1186\/gb-2011-12-4-220","article-title":"The cullin protein family","volume":"12","author":"Sarikas","year":"2011","journal-title":"Genome Biol."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"33","DOI":"10.1016\/j.gene.2015.08.064","article-title":"Distinct and overlapping functions of the cullin E3 ligase scaffolding proteins CUL4A and CUL4B","volume":"573","author":"Hannah","year":"2015","journal-title":"Gene"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"386","DOI":"10.1111\/j.1365-313X.2004.02302.x","article-title":"Molecular and functional characterization of Arabidopsis Cullin 3A","volume":"41","author":"Dieterle","year":"2005","journal-title":"Plant J."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"1024","DOI":"10.1016\/j.cell.2011.10.035","article-title":"The molecular basis of CRL4DDB2\/CSA ubiquitin ligase architecture, targeting, and activation","volume":"147","author":"Fischer","year":"2011","journal-title":"Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/j.ygeno.2013.01.004","article-title":"Transcriptome analysis of Chlamydomonas reinhardtii during the process of lipid accumulation","volume":"101","author":"Lv","year":"2013","journal-title":"Genomics"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1373","DOI":"10.1021\/pr400952z","article-title":"Integrated quantitative analysis of nitrogen stress response in Chlamydomonas reinhardtii using metabolite and protein profiling","volume":"13","author":"Wase","year":"2014","journal-title":"J. Proteome Res."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"3627","DOI":"10.1105\/tpc.111.087999","article-title":"Modulation of phototropic responsiveness in Arabidopsis through ubiquitination of phototropin 1 by the CUL3-Ring E3 ubiquitin ligase CRL3NPH3","volume":"23","author":"Roberts","year":"2011","journal-title":"Plant Cell"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"123","DOI":"10.1007\/s11105-017-1064-9","article-title":"Light regulates the RUBylation levels of individual Cullin proteins in Arabidopsis thaliana","volume":"36","author":"Christians","year":"2018","journal-title":"Plant Mol. Biol. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1007\/s11120-018-0580-2","article-title":"Changes in the photosynthetic apparatus and lipid droplet formation in Chlamydomonas reinhardtii under iron deficiency","volume":"139","author":"Devadasu","year":"2019","journal-title":"Photosynth. Res."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"65","DOI":"10.1074\/mcp.M112.021840","article-title":"The proteome of copper, iron, zinc, and manganese micronutrient deficiency in Chlamydomonas reinhardtii","volume":"12","author":"Hsieh","year":"2013","journal-title":"Mol. Cell. Proteom."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Beller, M., Sztalryd, C., Southall, N., Ming, B., and Oliver, B. (2008). CopI complex is a regulator of lipid homeostasis. PLoS Biol., 6.","DOI":"10.1371\/journal.pbio.0060292"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"611","DOI":"10.1111\/j.1365-313X.2004.02227.x","article-title":"Tandem inverted repeat system for selection of effective transgenic RNAi strains in Chlamydomonas","volume":"40","author":"Rohr","year":"2004","journal-title":"Plant J."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"387","DOI":"10.1146\/annurev-biochem-060410-105307","article-title":"Ubiquitin ligases and cell cycle control","volume":"82","author":"Teixeira","year":"2013","journal-title":"Annu. Rev. Biochem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"486","DOI":"10.1038\/70272","article-title":"CUL-2 is required for the G1-to-S-phase transition and mitotic chromosome condensation in Caenorhabditis elegans","volume":"1","author":"Feng","year":"1999","journal-title":"Nat. Cell Biol."},{"key":"ref_38","first-page":"1405","article-title":"Cul3 neddylation is crucial for gradual lipid droplet formation during adipogenesis","volume":"1864","author":"Dubiel","year":"2017","journal-title":"BBA Mol. Cell Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"2253","DOI":"10.1105\/tpc.112.107292","article-title":"Arabidopsis BPM proteins function as substrate adaptors to a cullin3-based E3 ligase to affect fatty acid metabolism in plants","volume":"25","author":"Chen","year":"2013","journal-title":"Plant Cell"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"973","DOI":"10.1016\/j.freeradbiomed.2011.11.038","article-title":"Activation of stress signaling pathways by electrophilic oxidized and nitrated lipids","volume":"52","author":"Kansanen","year":"2012","journal-title":"Free Radic. Biol. Med."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1437","DOI":"10.1105\/tpc.108.058891","article-title":"Arabidopsis DDB1-CUL4 associated factor1 forms a nuclear E3 ubiquitin ligase with DDB1 and CUL4 that is involved in multiple plant developmental processes","volume":"20","author":"Zhang","year":"2008","journal-title":"Plant Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"3814","DOI":"10.1038\/s41467-019-11772-y","article-title":"Structural basis of Cullin 2 RING E3 ligase regulation by the COP9 signalosome","volume":"10","author":"Faull","year":"2019","journal-title":"Nat. Commun."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1228","DOI":"10.1073\/pnas.87.3.1228","article-title":"High-frequency nuclear transformation of Chlamydomonas reinhardtii","volume":"87","author":"Kindle","year":"1990","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"691","DOI":"10.3389\/fpls.2018.00691","article-title":"Flagella-Associated WDR-Containing Protein CrFAP89 Regulates Growth and Lipid Accumulation in Chlamydomonas reinhardtii","volume":"9","author":"Luo","year":"2018","journal-title":"Front. Plant Sci."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4695\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:55:04Z","timestamp":1760162104000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/22\/9\/4695"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,29]]},"references-count":44,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["ijms22094695"],"URL":"https:\/\/doi.org\/10.3390\/ijms22094695","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,29]]}}}