ERα36, a variant of estrogen receptor α, is predominantly localized in mitochondria of human uterine smooth muscle and leiomyoma cells
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{"title"=>"ERα36, a variant of estrogen receptor α, is predominantly localized in mitochondria of human uterine smooth muscle and leiomyoma cells", "type"=>"journal", "authors"=>[{"first_name"=>"Yitang", "last_name"=>"Yan", "scopus_author_id"=>"57109614100"}, {"first_name"=>"Linda", "last_name"=>"Yu", "scopus_author_id"=>"24177980200"}, {"first_name"=>"Lysandra", "last_name"=>"Castro", "scopus_author_id"=>"36102300900"}, {"first_name"=>"Darlene", "last_name"=>"Dixon", "scopus_author_id"=>"7401748032"}], "year"=>2017, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203 (Electronic)\r1932-6203 (Linking)", "issn"=>"19326203", "pmid"=>"29020039", "sgr"=>"85031306919", "pui"=>"618698406", "scopus"=>"2-s2.0-85031306919", "doi"=>"10.1371/journal.pone.0186078"}, "id"=>"0eea5b59-03db-32a2-a45a-fd113637da31", "abstract"=>"ERalpha36 is a naturally occurring, membrane-associated, isoform of estrogen receptor alpha. The expression of ERalpha36 is due to alternative splicing and different promoter usage. ERalpha36 is a dominant-negative effector of ERalpha66-mediated transactivational activities and has the potential to trigger membrane-initiated mitogenic, nongenomic, estrogen signaling; however, the subcellular localization of ERalpha36 remains controversial. To determine the cellular localization of ERalpha36 in estrogen-responsive human uterine smooth muscle (ht-UtSMC) and leiomyoma (fibroid; ht-UtLM) cells, we conducted systematic confocal microscopy and subcellular fractionation analysis using ERalpha36 antibodies. With Image J colocalizaton analysis plugin, confocal images were analyzed to obtain a Pearson's Correlation Coefficient (PCC) to quantify signal colocalization of ERalpha36 with mitochondrial, endoplasmic reticulum, and cytoskeletal components in both cell lines. When cells were double-stained with an ERalpha36 antibody and a mitochondrial-specific dye, MitoTracker, the PCC for the two channel signals were both greater than 0.75, indicating strong correlation between ERalpha36 and mitochondrial signals in the two cell lines. A blocking peptide competition assay confirmed that the mitochondria-associated ERalpha36 signal detected by confocal analysis was specific for ERalpha36. In contrast, confocal images double-stained with an ERalpha36 antibody and endoplasmic reticulum or cytoskeletal markers, had PCCs that were all less than 0.4, indicating no or very weak signal correlation. Fractionation studies showed that ERalpha36 existed predominantly in membrane fractions, with minimal or undetected amounts in the cytosol, nuclear, chromatin, and cytoskeletal fractions. With isolated mitochondrial preparations, we confirmed that a known mitochondrial protein, prohibitin, was present in mitochondria, and by co-immunoprecipitation analysis that ERalpha36 was associated with prohibitin in ht-UtLM cells. The distinctive colocalization pattern of ERalpha36 with mitochondria in ht-UtSMC and ht-UtLM cells, and the association of ERalpha36 with a mitochondrial-specific protein suggest that ERalpha36 is localized primarily in mitochondria and may play a pivotal role in non-genomic signaling and mitochondrial functions.", "link"=>"http://www.mendeley.com/research/er%CE%B136-variant-estrogen-receptor-%CE%B1-predominantly-localized-mitochondria-human-uterine-smooth-muscle-l", "reader_count"=>5, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Doctoral Student"=>1, "Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>1, "Researcher"=>1, "Student > Doctoral Student"=>1, "Student > Master"=>1, "Student > Bachelor"=>1}, "reader_count_by_subject_area"=>{"Unspecified"=>1, "Biochemistry, Genetics and Molecular Biology"=>3, "Medicine and Dentistry"=>1}, "reader_count_by_subdiscipline"=>{"Medicine and Dentistry"=>{"Medicine and Dentistry"=>1}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}, "Unspecified"=>{"Unspecified"=>1}}, "group_count"=>0}

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