@article{fb45fafe038c43c0be1993583183d528,
title = "A chloroplast protein atlas reveals punctate structures and spatial organization of biosynthetic pathways",
abstract = "Chloroplasts are eukaryotic photosynthetic organelles that drive the global carbon cycle. Despite their importance, our understanding of their protein composition, function, and spatial organization remains limited. Here, we determined the localizations of 1,034 candidate chloroplast proteins using fluorescent protein tagging in the model alga Chlamydomonas reinhardtii. The localizations provide insights into the functions of poorly characterized proteins; identify novel components of nucleoids, plastoglobules, and the pyrenoid; and reveal widespread protein targeting to multiple compartments. We discovered and further characterized cellular organizational features, including eleven chloroplast punctate structures, cytosolic crescent structures, and unexpected spatial distributions of enzymes within the chloroplast. We also used machine learning to predict the localizations of other nuclear-encoded Chlamydomonas proteins. The strains and localization atlas developed here will serve as a resource to accelerate studies of chloroplast architecture and functions.",
keywords = "Chlamydomonas reinhardtii, chloroplast, dual targeting, fluorescent tagging, nucleoid, plastoglobule, protein localization, protein localization prediction, protein-protein interaction, pyrenoid",
author = "Lianyong Wang and Weronika Patena and {Van Baalen}, {Kelly A.} and Yihua Xie and Singer, {Emily R.} and Sophia Gavrilenko and Michelle Warren-Williams and Linqu Han and Harrigan, {Henry R.} and Hartz, {Linnea D.} and Vivian Chen and Ton, {Vinh T.N.P.} and Saw Kyin and Shwe, {Henry H.} and Cahn, {Matthew H.} and Wilson, {Alexandra T.} and Masayuki Onishi and Jianping Hu and Schnell, {Danny J.} and McWhite, {Claire D.} and Jonikas, {Martin C.}",
note = "Funding Information: We thank Princeton University Confocal Microscopy manager Gary Laevsky for instrumentation support; Princeton Institute for Computational Science and Engineering for research computing resources; Laurent Cournac, Ben Engel, Ursula Goodenough, Olivier Vallon, Christoph Benning, Ned Wingreen, Yonghua Li-Beisson, and present and former laboratory members for manuscript discussions; and Marie Bao, as part of Life Science Editors, for manuscript editing help. This material is based upon work supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research under award number DE-SC0020195; HHMI/Simons Foundation grant 55108535; and the Lewis-Sigler Scholars Fund. M.C.J. is a Howard Hughes Medical Institute Investigator. Conceptualization, L.W. and M.C.J.; gene cloning and confocal microscopy, L.W. K.A.V.B. Y.X. S.G. and H.R.H.; medium preparation, M.W.-W.; mass spectrometry analysis, S.K. H.H.S. L.W. E.R.S. and L.D.H.; prediction of protein localizations, C.D.M. and W.P.; prediction of protein structure, C.D.M. V.C. and V.T.N.P.T.; IF and immunoblotting assay, L.W.; characterization of CRISPR-Cas9 mutants, L.W. and M.O.; protein localization in tobacco, L.H. D.J.S. and J.H.; building the data/material-sharing website, M.H.C. W.P. L.W. and M.C.J. All authors analyzed the data. L.W. A.T.W. and M.C.J. wrote the manuscript, with input from all authors. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research. Funding Information: We thank Princeton University Confocal Microscopy manager Gary Laevsky for instrumentation support; Princeton Institute for Computational Science and Engineering for research computing resources; Laurent Cournac, Ben Engel, Ursula Goodenough, Olivier Vallon, Christoph Benning, Ned Wingreen, Yonghua Li-Beisson, and present and former laboratory members for manuscript discussions; and Marie Bao, as part of Life Science Editors, for manuscript editing help. This material is based upon work supported by the U.S. Department of Energy, Office of Science, Office of Biological and Environmental Research under award number DE-SC0020195 ; HHMI/Simons Foundation grant 55108535 ; and the Lewis-Sigler Scholars Fund . M.C.J. is a Howard Hughes Medical Institute Investigator. Publisher Copyright: {\textcopyright} 2023 The Authors",
year = "2023",
month = aug,
day = "3",
doi = "10.1016/j.cell.2023.06.008",
language = "English (US)",
volume = "186",
pages = "3499--3518.e14",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "16",
}