Localization of Lipid Raft Proteins to the Plasma Membrane Is a Major Function of the Phospholipid Transfer Protein Sec14
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{"title"=>"Localization of Lipid Raft Proteins to the Plasma Membrane Is a Major Function of the Phospholipid Transfer Protein Sec14", "type"=>"journal", "authors"=>[{"first_name"=>"Amy J.", "last_name"=>"Curwin", "scopus_author_id"=>"22134505500"}, {"first_name"=>"Marissa A.", "last_name"=>"LeBlanc", "scopus_author_id"=>"36673187500"}, {"first_name"=>"Gregory D.", "last_name"=>"Fairn", "scopus_author_id"=>"10039919500"}, {"first_name"=>"Christopher R.", "last_name"=>"McMaster", "scopus_author_id"=>"7004352587"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"368227209", "doi"=>"10.1371/journal.pone.0055388", "sgr"=>"84873865915", "scopus"=>"2-s2.0-84873865915", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"23383173"}, "id"=>"abcf17c0-f83e-3e65-a907-decef1943283", "abstract"=>"The Sec14 protein domain is a conserved tertiary structure that binds hydrophobic ligands. The Sec14 protein from Saccharomyces cerevisiae is essential with studies of S. cerevisiae Sec14 cellular function facilitated by a sole temperature sensitive allele, sec14(ts). The sec14(ts) allele encodes a protein with a point mutation resulting in a single amino acid change, Sec14(G266D). In this study results from a genome-wide genetic screen, and pharmacological data, provide evidence that the Sec14(G266D) protein is present at a reduced level compared to wild type Sec14 due to its being targeted to the proteosome. Increased expression of the sec14(ts) allele ameliorated growth arrest, but did not restore the defects in membrane accumulation or vesicular transport known to be defective in sec14(ts) cells. We determined that trafficking and localization of two well characterized lipid raft resident proteins, Pma1 and Fus-Mid-GFP, were aberrant in sec14(ts) cells. Localization of both lipid raft proteins was restored upon increased expression of the sec14(ts) allele. We suggest that a major function provided by Sec14 is trafficking and localization of lipid raft proteins.", "link"=>"http://www.mendeley.com/research/localization-lipid-raft-proteins-plasma-membrane-major-function-phospholipid-transfer-protein-sec14", "reader_count"=>35, "reader_count_by_academic_status"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Student > Doctoral Student"=>3, "Researcher"=>8, "Student > Ph. D. Student"=>8, "Student > Master"=>2, "Other"=>1, "Student > Bachelor"=>8, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_user_role"=>{"Unspecified"=>2, "Professor > Associate Professor"=>2, "Student > Doctoral Student"=>3, "Researcher"=>8, "Student > Ph. D. Student"=>8, "Student > Master"=>2, "Other"=>1, "Student > Bachelor"=>8, "Lecturer > Senior Lecturer"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>2, "Biochemistry, Genetics and Molecular Biology"=>5, "Medicine and Dentistry"=>1, "Agricultural and Biological Sciences"=>25, "Neuroscience"=>1, "Pharmacology, Toxicology and Pharmaceutical Science"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Neuroscience"=>{"Neuroscience"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>25}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>5}, "Unspecified"=>{"Unspecified"=>2}, "Pharmacology, Toxicology and Pharmaceutical Science"=>{"Pharmacology, Toxicology and Pharmaceutical Science"=>1}}, "reader_count_by_country"=>{"Netherlands"=>1, "Brazil"=>1, "Chile"=>1, "Germany"=>1}, "group_count"=>1}

Scopus | Further Information

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/498820"], "description"=>"<p>Yeast strains used in this study.</p>", "links"=>[], "tags"=>["strains"], "article_id"=>169342, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.t002", "stats"=>{"downloads"=>0, "page_views"=>4, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Yeast_strains_used_in_this_study_/169342", "title"=>"Yeast strains used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-19 16:16:34"}
  • {"files"=>["https://ndownloader.figshare.com/files/498746"], "description"=>"<p><b>Pma1 localization is defective in </b><b><i>sec14<sup>ts</sup></i></b><b> cells and restored by expression of Sec14<sup>G266D</sup>. </b><i>A, sec14<sup>ts</sup></i> cells expressing Fus-Mid-GFP and also containing empty vector, a vector expressing wild type Sec14 on a low copy plasmid, or Sec14<sup>G266D</sup> on a high copy (2 μ) plasmid, were grown at 25°C in 1% raffinose containing medium to mid-logarithmic phase. Cells were shifted to 37°C in pre-warmed 2% galactose containing medium for 3 hours. <i>B</i>, cells from <i>A</i> were quantified based on having only plasma membrane (PM) localization, only internal localization or both (vector n = 153, Sec14 n = 73, Sec14<sup>G266D</sup> n = 107) <i>C</i>, the wild type <i>SEC14</i> gene was replaced with the <i>sec14<sup>ts</sup></i> allele in a yeast strain expressing chimeric Pma1-RFP. The strain was transformed with either empty vector, a plasmid carried at low copy (ARS/CEN) containing wild type Sec14, and low and high copy (2 μ) plasmids containing Sec14<sup>G266D</sup>. Cells were grown at 25°C to mid-logarithmic phase and then transferred to 37°C for 16 hrs subsequent to determination Pma1-RFP localization by fluorescence microscopy.</p>", "links"=>[], "tags"=>["genetics and genomics", "molecular biology", "Computational biology", "Biochemistry"], "article_id"=>169260, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.g006", "stats"=>{"downloads"=>3, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Fus_Mid_GFP_and_/169260", "title"=>"Fus-Mid-GFP and", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:16:08"}
  • {"files"=>["https://ndownloader.figshare.com/files/498179"], "description"=>"<p>Serial dilutions of cells cultured at 25°C and then plated and grown at 25°C and 37°C for 3 days. <i>A</i>, wild type, <i>sec14<sup>ts</sup></i> and <i>sec14<sup>ts</sup> rpn4</i>Δ. <i>B</i>, The <i>sec14<sup>ts</sup></i> strain transformed with either empty vector, a plasmid carried at low copy (ARS/CEN) containing wild type Sec14, or a high copy (2 μ) plasmid containing Sec14<sup>G266D</sup>.</p>", "links"=>[], "tags"=>["sec14"], "article_id"=>168693, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.g001", "stats"=>{"downloads"=>0, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_Sec14_G266D_protein_can_provide_the_essential_function_of_Sec14_/168693", "title"=>"The Sec14 <sup>G266D</sup> protein can provide the essential function of Sec14.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:12:52"}
  • {"files"=>["https://ndownloader.figshare.com/files/498353"], "description"=>"<p><i>A</i>, the level of Sec14 and Sec14<sup>G266D</sup> in <i>sec14<sup>ts</sup> rpn4</i>Δ cells. <i>B</i>, the level of Sec14 and Sec14<sup>G266D</sup> in <i>sec14<sup>ts</sup></i> cells treated with MG132. Strains were transformed with plasmids expressing Sec14 or Sec14<sup>G266D</sup> containing an N-terminal T7 epitope tag and were grown to mid-logarithmic phase at 25°C, with a subset shifted to 37°C for 2 hours (A). For MG132 treatment cells were grown as before and shifted to 37°C in the presence of 100 μM MG132 for 2 hours. Cells were disrupted by three passes through a French press and unbroken cells removed by centrifugation. Protein extract was separated by SDS-PAGE, transferred to PVDF membrane, and western blots versus the T7 epitope were performed. Pgk1 was used as load control.</p>", "links"=>[], "tags"=>["levels", "regulated"], "article_id"=>168870, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.g003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_levels_of_Sec14_G266D_is_regulated_by_the_proteosome_/168870", "title"=>"The levels of Sec14<sup>G266D</sup> is regulated by the proteosome.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:13:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/498844"], "description"=>"<p><i>sec14<sup>ts</sup></i> suppressing gene deletions.</p>", "links"=>[], "tags"=>["suppressing"], "article_id"=>169361, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.t001", "stats"=>{"downloads"=>1, "page_views"=>5, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_sec14_ts_suppressing_gene_deletions_/169361", "title"=>"<i>sec14<sup>ts</sup></i> suppressing gene deletions.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-19 16:16:43"}
  • {"files"=>["https://ndownloader.figshare.com/files/498501"], "description"=>"<p>The <i>sec14<sup>ts</sup></i> strain transformed with either empty vector, a plasmid carried at low copy (ARS/CEN) containing wild type Sec14, or a high copy (2 μ) plasmid containing Sec14<sup>G266D</sup> were grown at 25°C to mid-logarithmic phase and then transferred to 37°C for 1 hr subsequent to determination of: <i>A</i>, invertase secretion (mean ± SE of three separate experiments performed in duplicate), <i>B</i>, or internal retention of Bgl2 at 2 and 16 hrs, similar results were seen at both time point with the 2 hr time point shown. <i>C</i>, the <i>sec14<sup>ts</sup></i> strain containing plasmid borne GFP-Snc1 was transformed with either empty vector, a plasmid carried at low copy (<i>ARS/CEN</i>) containing wild type Sec14, or a high copy (2 μ) plasmid containing Sec14<sup>G266D</sup>. Cells were grown at 25°C to mid-logarithmic phase and then transferred to 37°C for 2 hrs. The localization of GFP-Snc1 was determined by fluorescence microscopy in live cells. <i>D</i>, The strains were grown at 25°C to mid-logarithmic phase and then transferred to 37°C for 15 min prior to the addition of FM4-64. The trafficking of FM4-64 in live cells was visualized by fluorescence microscopy.</p>", "links"=>[], "tags"=>["vesicular", "trafficking", "pathways", "aberrant", "cells", "expressing"], "article_id"=>169014, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.g004", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Known_vesicular_trafficking_pathways_are_still_aberrant_in_growing_cells_expressing_Sec14_G266D_/169014", "title"=>"Known vesicular trafficking pathways are still aberrant in growing cells expressing Sec14<sup>G266D</sup>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:14:47"}
  • {"files"=>["https://ndownloader.figshare.com/files/498639"], "description"=>"<p>Wild type cells and <i>sec14<sup>ts</sup></i> cells containing empty vector, a vector expressing wild type Sec14 on a low copy plasmid, or Sec14<sup>G266D</sup> on a high copy (2 μ) plasmid, were grown at 25°C to mid-logarithmic phase and an aliquot transferred to 37°C for 1 hr followed by incubation in 1.5% KMnO<sub>4</sub>, 1% sodium periodate, and then 1% NH<sub>4</sub>Cl subsequent to embedding and viewing by transmission electron microscopy.</p>", "links"=>[], "tags"=>["accumulate", "cells", "expressing"], "article_id"=>169158, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.g005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Membranes_accumulate_in_growing_cells_expressing_Sec14_G266D_/169158", "title"=>"Membranes accumulate in growing cells expressing Sec14<sup>G266D</sup>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:15:35"}
  • {"files"=>["https://ndownloader.figshare.com/files/498272"], "description"=>"<p><i>A</i>, the <i>sec14<sup>ts</sup></i> cells were transformed with a plasmid expressing Sec14 containing an N-terminal T7 epitope, untagged Sec14, or empty vector. <i>SEC14</i> expression was driven by the constitutive <i>GPD1</i> promoter. Cells were grown in solution at 25°C to mid-logarithmic phase, and serial dilutions of identical numbers of cells were spotted onto plates and incubated at 37°C for two days. <i>B</i>, cells expressing T7-Sec14 or Sec14<sup>G266D</sup> were grown to mid-logarithmic phase at 25°C, with a subset shifted to 37°C for 2 hrs. Cells were disrupted by three passes through a French press and membranes were separated from soluble proteins by differential centrifugation. Proteins in each fraction were separated by SDS-PAGE, transferred to PVDF membrane, and western blots were performed. In the blots shown 10 fold more protein extract was loaded in each Sec14<sup>G266D</sup> lane compared to extracts containing wild type Sec14 for blots versus the T7 epitope due to protein expression level differences.</p>", "links"=>[], "tags"=>["sec14"], "article_id"=>168799, "categories"=>["Molecular Biology", "Biochemistry", "Genetics", "Biological Sciences"], "users"=>["Amy J. Curwin", "Marissa A. LeBlanc", "Gregory D. Fairn", "Christopher R. McMaster"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055388.g002", "stats"=>{"downloads"=>1, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_temperature_on_Sec14_and_Sec14_G266D_protein_levels_/168799", "title"=>"Effect of temperature on Sec14 and Sec14 <sup>G266D</sup> protein levels.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-19 16:13:30"}

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Relative Metric

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