气候变化领域集成服务门户(CCPortal): Widespread polycistronic gene expression in green algae

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DOI	10.1073/pnas.2017714118
	Widespread polycistronic gene expression in green algae
	Gallaher S.D.; Craig R.J.; Ganesan I.; Purvine S.O.; McCorkle S.R.; Grimwood J.; Strenkert D.; Davidi L.; Roth M.S.; Jeffers T.L.; Lipton M.S.; Niyogi K.K.; Schmutz J.; Theg S.M.; Blaby-Haas C.E.; Merchant S.S.
发表日期	2021
ISSN	00278424
卷号	118 期号:7
英文摘要	Polycistronic gene expression, common in prokaryotes, was thought to be extremely rare in eukaryotes. The development of long-read sequencing of full-length transcript isomers (Iso-Seq) has facilitated a reexamination of that dogma. Using Iso-Seq, we discovered hundreds of examples of polycistronic expression of nuclear genes in two divergent species of green algae: Chlamydomonas reinhardtii and Chromochloris zofingiensis. Here, we employ a range of independent approaches to validate that multiple proteins are translated from a common transcript for hundreds of loci. A chromatin immunoprecipitation analysis using trimethylation of lysine 4 on histone H3 marks confirmed that transcription begins exclusively at the upstream gene. Quantification of polyadenylated [poly(A)] tails and poly(A) signal sequences confirmed that transcription ends exclusively after the downstream gene. Coexpression analysis found nearly perfect correlation for open reading frames (ORFs) within polycistronic loci, consistent with expression in a shared transcript. For many polycistronic loci, terminal peptides from both ORFs were identified from proteomics datasets, consistent with independent translation. Synthetic polycistronic gene pairs were transcribed and translated in vitro to recapitulate the production of two distinct proteins from a common transcript. The relative abundance of these two proteins can be modified by altering the Kozak-like sequence of the upstream gene. Replacement of the ORFs with selectable markers or reporters allows production of such heterologous proteins, speaking to utility in synthetic biology approaches. Conservation of a significant number of polycistronic gene pairs between C. reinhardtii, C. zofingiensis, and five other species suggests that this mechanism may be evolutionarily ancient and biologically important in the green algal lineage. © 2021 National Academy of Sciences. All rights reserved.
英文关键词	Transcriptome \| bicistronic \| dicistronic \| leaky ribosome scanning \| uORFs
语种	英语
scopus关键词	algal protein; histone H3; lysine; Article; chromatin immunoprecipitation; controlled study; downstream processing; gene expression; gene locus; genetic correlation; genetic transcription; green alga; in vitro study; nonhuman; open reading frame; priority journal; proteomics; RNA translation; synthetic biology; validation process
来源期刊	Proceedings of the National Academy of Sciences of the United States of America
文献类型	期刊论文
条目标识符	http://gcip.llas.ac.cn/handle/2XKMVOVA/180641
作者单位	UCLA DOE Institute for Genomics and Proteomics, University of California, Los Angeles, CA 90095, United States; Department of Chemistry and Biochemistry, University of California, Los Angeles, CA 90095, United States; Joint Genome Institute, United States Department of Energy, Berkeley, CA 94720, United States; Institute of Evolutionary Biology, School of Biological Sciences, University of Edinburgh, Edinburgh, EH9 3FL, United Kingdom; Department of Plant Biology, University of California, Davis, CA 95616, United States; Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99354, United States; Biology Department, Brookhaven National Laboratory, Upton, NY 11973, United States; HudsonAlpha Genome Sequencing Center, HudsonAlpha Institute for Biotechnology, Huntsville, AL 35806, United States; Department of Plant and Microbial Biology, University of California, Berkeley, CA 94720, United States; Division of Molecular Biophysics and Integrated Bioimaging, La...
推荐引用方式 GB/T 7714	Gallaher S.D.,Craig R.J.,Ganesan I.,et al. Widespread polycistronic gene expression in green algae[J],2021,118(7).
APA	Gallaher S.D..,Craig R.J..,Ganesan I..,Purvine S.O..,McCorkle S.R..,...&Merchant S.S..(2021).Widespread polycistronic gene expression in green algae.Proceedings of the National Academy of Sciences of the United States of America,118(7).
MLA	Gallaher S.D.,et al."Widespread polycistronic gene expression in green algae".Proceedings of the National Academy of Sciences of the United States of America 118.7(2021).