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  1. AU="Lopez-Paz, Cristina"
  2. AU="Rattenbacher, Bernd"
  3. AU="Smith, K. L"
  4. AU="Nasr, Suhayl J"
  5. AU="Wade, Nekeithia S"
  6. AU="Carabellese, Felice"
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  11. AU="Catanesi, M. G."
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  16. AU="Gallagher, Julia E"
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  38. AU="Georgel, Philippe"
  39. AU="Gruenewald, Leon D"
  40. AU="Charron, Morgane"
  41. AU="Leona S. Alizadeh" AU="Leona S. Alizadeh"
  42. AU="Soriano, Stéphane"
  43. AU="Lin, Pao-Yen"
  44. AU="Mudali, Gayathri"
  45. AU="McElveen, John T"
  46. AU="Kraimps, Jean-Louis"
  47. AU="Patel, Sheila K"
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  49. AU="Langley, Jonathan"
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  1. Artikel ; Online: Subscaling of a cytosolic RNA binding protein governs cell size homeostasis in the multiple fission alga Chlamydomonas.

    Liu, Dianyi / Lopez-Paz, Cristina / Li, Yubing / Zhuang, Xiaohong / Umen, James

    PLoS genetics

    2024  Band 20, Heft 3, Seite(n) e1010503

    Abstract: Coordination of growth and division in eukaryotic cells is essential for populations of proliferating cells to maintain size homeostasis, but the underlying mechanisms that govern cell size have only been investigated in a few taxa. The green alga ... ...

    Abstract Coordination of growth and division in eukaryotic cells is essential for populations of proliferating cells to maintain size homeostasis, but the underlying mechanisms that govern cell size have only been investigated in a few taxa. The green alga Chlamydomonas reinhardtii (Chlamydomonas) proliferates using a multiple fission cell cycle that involves a long G1 phase followed by a rapid series of successive S and M phases (S/M) that produces 2n daughter cells. Two control points show cell-size dependence: the Commitment control point in mid-G1 phase requires the attainment of a minimum size to enable at least one mitotic division during S/M, and the S/M control point where mother cell size governs cell division number (n), ensuring that daughter distributions are uniform. tny1 mutants pass Commitment at a smaller size than wild type and undergo extra divisions during S/M phase to produce small daughters, indicating that TNY1 functions to inhibit size-dependent cell cycle progression. TNY1 encodes a cytosolic hnRNP A-related RNA binding protein and is produced once per cell cycle during S/M phase where it is apportioned to daughter cells, and then remains at constant absolute abundance as cells grow, a property known as subscaling. Altering the dosage of TNY1 in heterozygous diploids or through mis-expression increased Commitment cell size and daughter cell size, indicating that TNY1 is a limiting factor for both size control points. Epistasis placed TNY1 function upstream of the retinoblastoma tumor suppressor complex (RBC) and one of its regulators, Cyclin-Dependent Kinase G1 (CDKG1). Moreover, CDKG1 protein and mRNA were found to over-accumulate in tny1 cells suggesting that CDKG1 may be a direct target of repression by TNY1. Our data expand the potential roles of subscaling proteins outside the nucleus and imply a control mechanism that ties TNY1 accumulation to pre-division mother cell size.
    Mesh-Begriff(e) Chlamydomonas/metabolism ; Cell Cycle/genetics ; Cell Division ; Cyclin-Dependent Kinases/genetics ; RNA-Binding Proteins/genetics ; Cell Size
    Chemische Substanzen Cyclin-Dependent Kinases (EC 2.7.11.22) ; RNA-Binding Proteins
    Sprache Englisch
    Erscheinungsdatum 2024-03-18
    Erscheinungsland United States
    Dokumenttyp Journal Article
    ZDB-ID 2186725-2
    ISSN 1553-7404 ; 1553-7390
    ISSN (online) 1553-7404
    ISSN 1553-7390
    DOI 10.1371/journal.pgen.1010503
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  2. Artikel ; Online: Identification of Chlamydomonas reinhardtii endogenous genic flanking sequences for improved transgene expression.

    López-Paz, Cristina / Liu, Dianyi / Geng, Sa / Umen, James G

    The Plant journal : for cell and molecular biology

    2017  Band 92, Heft 6, Seite(n) 1232–1244

    Abstract: Chlamydomonas reinhardtii is a unicellular green alga that has attracted interest due to its potential biotechnological applications, and as a model for algal biofuel and energy metabolism. Despite all the advantages that this unicellular alga offers, ... ...

    Abstract Chlamydomonas reinhardtii is a unicellular green alga that has attracted interest due to its potential biotechnological applications, and as a model for algal biofuel and energy metabolism. Despite all the advantages that this unicellular alga offers, poor and inconsistent expression of nuclear transgenes remains an obstacle for basic and applied research. We used a data-mining strategy to identify highly expressed genes in Chlamydomonas whose flanking sequences were tested for the ability to drive heterologous nuclear transgene expression. Candidates identified in this search included two ribosomal protein genes, RPL35a and RPL23, and ferredoxin, FDX1, whose flanking regions including promoters, terminators and untranslated sequences could drive stable luciferase transgene expression to significantly higher levels than the commonly used Hsp70A-RBCS2 (AR) hybrid promoter/terminator sequences. The RPL23 flanking sequences were further tested using the zeocin resistance gene sh-ble as a reporter in monocistronic and dicistronic constructs, and consistently yielded higher numbers of zeocin-resistant transformants and higher levels of resistance than AR- or PSAD-based vectors. Chlamydomonas RPL23 sequences also enabled transgene expression in Volvox carteri. Our study provides an additional benchmark for strong constitutive expression of transgenes in Chlamydomonas, and develops a general approach for identifying flanking sequences that can be used to drive transgene expression for any organism where transcriptome data are available.
    Mesh-Begriff(e) 3' Flanking Region/genetics ; 5' Flanking Region/genetics ; Cell Nucleus/metabolism ; Chlamydomonas reinhardtii/genetics ; Gene Expression ; Genetic Vectors/genetics ; Luciferases/genetics ; Promoter Regions, Genetic/genetics ; Terminator Regions, Genetic/genetics ; Transgenes ; Untranslated Regions/genetics ; Volvox/genetics
    Chemische Substanzen Untranslated Regions ; Luciferases (EC 1.13.12.-)
    Sprache Englisch
    Erscheinungsdatum 2017-12
    Erscheinungsland England
    Dokumenttyp Journal Article
    ZDB-ID 1088037-9
    ISSN 1365-313X ; 0960-7412
    ISSN (online) 1365-313X
    ISSN 0960-7412
    DOI 10.1111/tpj.13731
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  3. Artikel: Identification of Chlamydomonas reinhardtii endogenous genic flanking sequences for improved transgene expression

    López‐Paz, Cristina / Liu, Dianyi / Geng, Sa / Umen, James G

    plant journal. 2017 Dec., v. 92, no. 6

    2017  

    Abstract: Chlamydomonas reinhardtii is a unicellular green alga that has attracted interest due to its potential biotechnological applications, and as a model for algal biofuel and energy metabolism. Despite all the advantages that this unicellular alga offers, ... ...

    Abstract Chlamydomonas reinhardtii is a unicellular green alga that has attracted interest due to its potential biotechnological applications, and as a model for algal biofuel and energy metabolism. Despite all the advantages that this unicellular alga offers, poor and inconsistent expression of nuclear transgenes remains an obstacle for basic and applied research. We used a data‐mining strategy to identify highly expressed genes in Chlamydomonas whose flanking sequences were tested for the ability to drive heterologous nuclear transgene expression. Candidates identified in this search included two ribosomal protein genes, RPL35a and RPL23, and ferredoxin, FDX1, whose flanking regions including promoters, terminators and untranslated sequences could drive stable luciferase transgene expression to significantly higher levels than the commonly used Hsp70A‐RBCS2 (AR) hybrid promoter/terminator sequences. The RPL23 flanking sequences were further tested using the zeocin resistance gene sh‐ble as a reporter in monocistronic and dicistronic constructs, and consistently yielded higher numbers of zeocin‐resistant transformants and higher levels of resistance than AR‐ or PSAD‐based vectors. Chlamydomonas RPL23 sequences also enabled transgene expression in Volvox carteri. Our study provides an additional benchmark for strong constitutive expression of transgenes in Chlamydomonas, and develops a general approach for identifying flanking sequences that can be used to drive transgene expression for any organism where transcriptome data are available.
    Schlagwörter Chlamydomonas reinhardtii ; Volvox ; algae ; applied research ; biofuels ; energy metabolism ; gene expression ; luciferase ; models ; promoter regions ; resistance genes ; ribosomal proteins ; terminator regions ; transcriptome ; transgenes
    Sprache Englisch
    Erscheinungsverlauf 2017-12
    Umfang p. 1232-1244.
    Erscheinungsort John Wiley & Sons, Ltd
    Dokumenttyp Artikel
    Anmerkung JOURNAL ARTICLE
    ZDB-ID 1088037-9
    ISSN 1365-313X ; 0960-7412
    ISSN (online) 1365-313X
    ISSN 0960-7412
    DOI 10.1111/tpj.13731
    Datenquelle NAL Katalog (AGRICOLA)

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  4. Artikel ; Online: A new class of cyclin dependent kinase in Chlamydomonas is required for coupling cell size to cell division.

    Li, Yubing / Liu, Dianyi / López-Paz, Cristina / Olson, Bradley Jsc / Umen, James G

    eLife

    2016  Band 5, Seite(n) e10767

    Abstract: Proliferating cells actively control their size by mechanisms that are poorly understood. The unicellular green alga Chlamydomonas reinhardtii divides by multiple fission, wherein a 'counting' mechanism couples mother cell-size to cell division number ... ...

    Abstract Proliferating cells actively control their size by mechanisms that are poorly understood. The unicellular green alga Chlamydomonas reinhardtii divides by multiple fission, wherein a 'counting' mechanism couples mother cell-size to cell division number allowing production of uniform-sized daughters. We identified a sizer protein, CDKG1, that acts through the retinoblastoma (RB) tumor suppressor pathway as a D-cyclin-dependent RB kinase to regulate mitotic counting. Loss of CDKG1 leads to fewer mitotic divisions and large daughters, while mis-expression of CDKG1 causes supernumerous mitotic divisions and small daughters. The concentration of nuclear-localized CDKG1 in pre-mitotic cells is set by mother cell size, and its progressive dilution and degradation with each round of cell division may provide a link between mother cell-size and mitotic division number. Cell-size-dependent accumulation of limiting cell cycle regulators such as CDKG1 is a potentially general mechanism for size control.
    Mesh-Begriff(e) Cell Division ; Cell Size ; Chlamydomonas reinhardtii/enzymology ; Chlamydomonas reinhardtii/genetics ; Cyclin-Dependent Kinases/genetics ; Cyclin-Dependent Kinases/metabolism ; Gene Expression ; Gene Knockout Techniques ; Signal Transduction
    Chemische Substanzen Cyclin-Dependent Kinases (EC 2.7.11.22)
    Sprache Englisch
    Erscheinungsdatum 2016-03-25
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2687154-3
    ISSN 2050-084X ; 2050-084X
    ISSN (online) 2050-084X
    ISSN 2050-084X
    DOI 10.7554/eLife.10767
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  5. Artikel ; Online: Defects in a new class of sulfate/anion transporter link sulfur acclimation responses to intracellular glutathione levels and cell cycle control.

    Fang, Su-Chiung / Chung, Chin-Lin / Chen, Chun-Han / Lopez-Paz, Cristina / Umen, James G

    Plant physiology

    2014  Band 166, Heft 4, Seite(n) 1852–1868

    Abstract: We previously identified a mutation, suppressor of mating type locus3 15-1 (smt15-1), that partially suppresses the cell cycle defects caused by loss of the retinoblastoma tumor suppressor-related protein encoded by the MAT3 gene in Chlamydomonas ... ...

    Abstract We previously identified a mutation, suppressor of mating type locus3 15-1 (smt15-1), that partially suppresses the cell cycle defects caused by loss of the retinoblastoma tumor suppressor-related protein encoded by the MAT3 gene in Chlamydomonas reinhardtii. smt15-1 single mutants were also found to have a cell cycle defect leading to a small-cell phenotype. SMT15 belongs to a previously uncharacterized subfamily of putative membrane-localized sulfate/anion transporters that contain a sulfate transporter domain and are found in a widely distributed subset of eukaryotes and bacteria. Although we observed that smt15-1 has a defect in acclimation to sulfur-limited growth conditions, sulfur acclimation (sac) mutants, which are more severely defective for acclimation to sulfur limitation, do not have cell cycle defects and cannot suppress mat3. Moreover, we found that smt15-1, but not sac mutants, overaccumulates glutathione. In wild-type cells, glutathione fluctuated during the cell cycle, with highest levels in mid G1 phase and lower levels during S and M phases, while in smt15-1, glutathione levels remained elevated during S and M. In addition to increased total glutathione levels, smt15-1 cells had an increased reduced-to-oxidized glutathione redox ratio throughout the cell cycle. These data suggest a role for SMT15 in maintaining glutathione homeostasis that impacts the cell cycle and sulfur acclimation responses.
    Mesh-Begriff(e) Acclimatization ; Algal Proteins/genetics ; Algal Proteins/metabolism ; Anion Transport Proteins/genetics ; Anion Transport Proteins/metabolism ; Anions/metabolism ; Base Sequence ; Cell Cycle ; Cell Cycle Checkpoints ; Chlamydomonas reinhardtii/genetics ; Chlamydomonas reinhardtii/physiology ; Cytoplasm/metabolism ; Glutathione/metabolism ; High-Throughput Nucleotide Sequencing ; Molecular Sequence Data ; Mutation ; Phylogeny ; Sequence Analysis, RNA ; Sulfates/metabolism ; Sulfur/metabolism
    Chemische Substanzen Algal Proteins ; Anion Transport Proteins ; Anions ; Sulfates ; Sulfur (70FD1KFU70) ; Glutathione (GAN16C9B8O)
    Sprache Englisch
    Erscheinungsdatum 2014-10-31
    Erscheinungsland United States
    Dokumenttyp Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 208914-2
    ISSN 1532-2548 ; 0032-0889
    ISSN (online) 1532-2548
    ISSN 0032-0889
    DOI 10.1104/pp.114.251009
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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  6. Artikel ; Online: Maize AKINbetagamma dimerizes through the KIS/CBM domain and assembles into SnRK1 complexes.

    López-Paz, Cristina / Vilela, Belmiro / Riera, Marta / Pagès, Montserrat / Lumbreras, Victoria

    FEBS letters

    2009  Band 583, Heft 12, Seite(n) 1887–1894

    Abstract: The SNF1/AMPK/SnRK1 complex is an intracellular energy sensor composed of three types of subunits: the SnRK1 kinase and two regulatory, non-catalytic subunits (designated beta and gamma). We have previously described an atypical plant gamma-subunit, ... ...

    Abstract The SNF1/AMPK/SnRK1 complex is an intracellular energy sensor composed of three types of subunits: the SnRK1 kinase and two regulatory, non-catalytic subunits (designated beta and gamma). We have previously described an atypical plant gamma-subunit, AKINbetagamma, which contains an N-terminal tail similar to the so-called KIS domain normally present in beta-subunits. However, it is not known whether AKINbetagamma normally associates with endogenous SnRK1 complexes in vivo, nor how its unique domain structure might contribute to SnRK1 function. Here, we present evidence that maize AKINbetagamma is an integral component of active SnRK1 complexes in plant cells. Using complementary methodological approaches, we also show that AKINbetagamma associates through homomeric interactions mediated by both, the gamma- and, unexpectedly, the KIS/CBM domain.
    Mesh-Begriff(e) Amino Acid Sequence ; Arabidopsis ; Bacterial Proteins/genetics ; Bacterial Proteins/metabolism ; Carbohydrate Metabolism ; Cells, Cultured ; Dimerization ; Luminescent Proteins/genetics ; Luminescent Proteins/metabolism ; Molecular Sequence Data ; Onions ; Plant Proteins/chemistry ; Plant Proteins/genetics ; Plant Proteins/metabolism ; Plants, Genetically Modified ; Protein Structure, Tertiary ; Protein Subunits ; Protein-Serine-Threonine Kinases/chemistry ; Protein-Serine-Threonine Kinases/genetics ; Protein-Serine-Threonine Kinases/metabolism ; Recombinant Fusion Proteins/chemistry ; Recombinant Fusion Proteins/genetics ; Recombinant Fusion Proteins/metabolism ; Sequence Homology, Amino Acid ; Zea mays/enzymology ; Zea mays/genetics
    Chemische Substanzen Bacterial Proteins ; Luminescent Proteins ; Plant Proteins ; Protein Subunits ; Recombinant Fusion Proteins ; yellow fluorescent protein, Bacteria ; SNF1-related protein kinases (EC 2.7.1.-) ; Protein-Serine-Threonine Kinases (EC 2.7.11.1)
    Sprache Englisch
    Erscheinungsdatum 2009-06-18
    Erscheinungsland England
    Dokumenttyp Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 212746-5
    ISSN 1873-3468 ; 0014-5793
    ISSN (online) 1873-3468
    ISSN 0014-5793
    DOI 10.1016/j.febslet.2009.05.022
    Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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