A Polymorphic 3’UTR Element in ATP1B1 Regulates Alternative Polyadenylation and Is Associated with Blood Pressure
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{"title"=>"A Polymorphic 3'UTR Element in ATP1B1 Regulates Alternative Polyadenylation and Is Associated with Blood Pressure", "type"=>"journal", "authors"=>[{"first_name"=>"Megana K.", "last_name"=>"Prasad", "scopus_author_id"=>"56349567100"}, {"first_name"=>"Kavita", "last_name"=>"Bhalla", "scopus_author_id"=>"54404693800"}, {"first_name"=>"Zhen Hua", "last_name"=>"Pan", "scopus_author_id"=>"55467956900"}, {"first_name"=>"Jeffrey R.", "last_name"=>"O'Connell", "scopus_author_id"=>"7202443550"}, {"first_name"=>"Alan B.", "last_name"=>"Weder", "scopus_author_id"=>"7005293856"}, {"first_name"=>"Aravinda", "last_name"=>"Chakravarti", "scopus_author_id"=>"35355137200"}, {"first_name"=>"Bin", "last_name"=>"Tian", "scopus_author_id"=>"7201678245"}, {"first_name"=>"Yen Pei C", "last_name"=>"Chang", "scopus_author_id"=>"8158751700"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"issn"=>"19326203", "pui"=>"369964010", "sgr"=>"84885062513", "doi"=>"10.1371/journal.pone.0076290", "scopus"=>"2-s2.0-84885062513", "isbn"=>"1932-6203 (Electronic)\\r1932-6203 (Linking)", "pmid"=>"24098465"}, "id"=>"25db7888-d1fa-3e05-b5e8-550d9cbf487b", "abstract"=>"Although variants in many genes have previously been shown to be associated with blood pressure (BP) levels, the molecular mechanism underlying these associations are mostly unknown. We identified a multi-allelic T-rich sequence (TRS) in the 3'UTR of ATP1B1 that varies in length and sequence composition (T22-27 and T12GT 3GT6). The 3'UTR of ATP1B1 contains 2 functional polyadenylation signals and the TRS is downstream of the proximal polyadenylation site (A2). Therefore, we hypothesized that alleles of this TRS might influence ATP1B1 expression by regulating alternative polyadenylation. In vitro, the T12GT 3GT6 allele increases polyadenylation at the A2 polyadenylation site as compared to the T23 allele. Consistent with our hypothesis, the relative abundance of the A2-polyadenylated ATP1B1 mRNA was higher in human kidneys with at least one copy of the T12GT 3GT6 allele than in those lacking this allele. The T12GT 3GT6 allele is also associated with higher systolic BP (beta = 3.3 mmHg, p = 0.014) and diastolic BP (beta = 2.4 mmHg, p = 0.003) in a European-American population. Therefore, we have identified a novel multi-allelic TRS in the 3'UTR of ATP1B1 that is associated with higher BP and may mediate its effect by regulating the polyadenylation of the ATP1B1 mRNA.", "link"=>"http://www.mendeley.com/research/polymorphic-3utr-element-atp1b1-regulates-alternative-polyadenylation-associated-blood-pressure", "reader_count"=>11, "reader_count_by_academic_status"=>{"Professor > Associate Professor"=>2, "Researcher"=>5, "Student > Ph. D. Student"=>3, "Student > Master"=>1}, "reader_count_by_user_role"=>{"Professor > Associate Professor"=>2, "Researcher"=>5, "Student > Ph. D. Student"=>3, "Student > Master"=>1}, "reader_count_by_subject_area"=>{"Biochemistry, Genetics and Molecular Biology"=>3, "Agricultural and Biological Sciences"=>8}, "reader_count_by_subdiscipline"=>{"Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>8}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>3}}, "reader_count_by_country"=>{"Germany"=>1}, "group_count"=>0}

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Figshare

  • {"files"=>["https://ndownloader.figshare.com/files/1221311"], "description"=>"<p>A) Genomic architecture of the <i>ATP1B1</i> gene. The red box highlights the 3’UTR, which is enlarged and shown below. TGA: translational stop codon, *: G/A: SNP rs12079745, A2: A2 polyadenylation signal, TRS: T-rich element, A5: A5 polyadenylation signal. B) Representative Sanger sequencing chromatograms of the T<sub>23</sub> and T <sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> alleles. Alleles are named based on the portion of the TRS that is unique to each allele (denoted by a horizontal bar). C) Sequence alignment of the TRS across mammals. The A2 polyadenylation signal and the TRS are highlighted in red and green respectively. D) Aligned are the 2a consensus sequence of the CstF binding site [<a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0076290#B38\" target=\"_blank\">38</a>] and the sequences of the TRS with the T<sub>23</sub> and T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> alleles. Asterisks mark the nucleotides in the TRS that deviate from the consensus CstF binding sequence. Y = C/T, N = A/C/G/T.</p>", "links"=>[], "tags"=>["genotyping", "t-rich"], "article_id"=>812220, "categories"=>["Biological Sciences"], "users"=>["Megana K. Prasad", "Kavita Bhalla", "Zhen Hua Pan", "Jeffrey R. O’Connell", "Alan B. Weder", "Aravinda Chakravarti", "Bin Tian", "Yen-Pei C. Chang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076290.g001"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Structure_and_genotyping_of_the_T_rich_sequence_in_ATP1B1_/812220", "title"=>"Structure and genotyping of the T-rich sequence in <i>ATP1B1</i>.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-01 02:12:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1221313"], "description"=>"<p>A) Schematic of the pRIG vector. MCS: multiple cloning site, where the tested sequences were inserted, pCMV: cytomegalovirus minimal promoter, RFP: red fluorescent protein, IRES: internal ribosomal entry site, eGFP: enhanced green fluorescent protein, SV40 pA: SV40 polyadenylation signal. B) Dot plots from FACS analysis of the expression of RFP and eGFP in HEK293T cells transfected with the T<sub>23</sub> and T <sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> alleles in the pRIG vector. Each dot represents a cell and the X- and Y-axes are the fluorescence intensity of eGFP and RFP, respectively. C) The ratio of RFP to eGFP expression in pRIG with no insert, T<sub>23</sub>, and T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub>.</p>", "links"=>[], "tags"=>["allele", "polyadenylation", "a2"], "article_id"=>812222, "categories"=>["Biological Sciences"], "users"=>["Megana K. Prasad", "Kavita Bhalla", "Zhen Hua Pan", "Jeffrey R. O’Connell", "Alan B. Weder", "Aravinda Chakravarti", "Bin Tian", "Yen-Pei C. Chang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076290.g002"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_T_12_GT_3_GT_6_allele_increases_polyadenylation_at_the_A2_polyadenylation_site_/812222", "title"=>"The T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele increases polyadenylation at the A2 polyadenylation site.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-01 02:12:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1221315"], "description"=>"<p>A) Schematic of the primer design used to distinguish between the A2- and A5- polyadenylated mRNAs. The A2-R primer had a 5’ T<sub>15</sub> tail to selectively amplify A2-polyadenylated mRNA. Total <i>ATP1B1</i> mRNA was measured using primers that amplified across the exon 5/exon 6 junction. B) Real-time PCR of mRNA from human kidneys. The A2-polyadenylated transcript was quantified relative to total <i>ATP1B1</i> mRNA. 0 and 1 indicate subjects without the T <sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele and with 1 copy of the T <sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele, respectively. Error bars represent standard errors. *<i>p</i> < 0.0001. C) Real-time PCR of mRNA from human lymphocytes. Expression of the A2- and A5-polyadenylated mRNAs relative to total <i>ATP1B1</i> mRNA in lymphocytes from subjects with 0 or at least 1 copy of the T <sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele. *<i>p</i> = 0.0001 between the A5-polyadenylated transcript levels in the 2 groups. Data represent the average across samples from each genotype group with standard error. D) Expression of ATP1B1 protein in lymphocytes from individuals with 0 or at least 1 copy of the T <sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele. Tubulin was used as a loading control. The graph quantifies the average level of ATP1B1 protein relative to tubulin with standard error. *<i>p</i> = 0.05.</p>", "links"=>[], "tags"=>["allele", "regulates", "polyadenylation", "mrna"], "article_id"=>812224, "categories"=>["Biological Sciences"], "users"=>["Megana K. Prasad", "Kavita Bhalla", "Zhen Hua Pan", "Jeffrey R. O’Connell", "Alan B. Weder", "Aravinda Chakravarti", "Bin Tian", "Yen-Pei C. Chang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076290.g003"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_The_T_12_GT_3_GT_6_allele_regulates_the_polyadenylation_of_ATP1B1_mRNA_in_human_tissues_/812224", "title"=>"The T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele regulates the polyadenylation of <i>ATP1B1</i> mRNA in human tissues", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-10-01 02:12:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/1221316", "https://ndownloader.figshare.com/files/1221317", "https://ndownloader.figshare.com/files/1221318", "https://ndownloader.figshare.com/files/1221319", "https://ndownloader.figshare.com/files/1221320"], "description"=>"<div><p>Although variants in many genes have previously been shown to be associated with blood pressure (BP) levels, the molecular mechanism underlying these associations are mostly unknown. We identified a multi-allelic T-rich sequence (TRS) in the 3’UTR of <i>ATP1B1</i> that varies in length and sequence composition (T<sub>22-27</sub> and T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub>). The 3’UTR of <i>ATP1B1</i> contains 2 functional polyadenylation signals and the TRS is downstream of the proximal polyadenylation site (A2). Therefore, we hypothesized that alleles of this TRS might influence <i>ATP1B1</i> expression by regulating alternative polyadenylation. <i>In vitro</i>, the T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele increases polyadenylation at the A2 polyadenylation site as compared to the T<sub>23</sub> allele. Consistent with our hypothesis, the relative abundance of the A2-polyadenylated <i>ATP1B1</i> mRNA was higher in human kidneys with at least one copy of the T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele than in those lacking this allele. The T<sub>12</sub>GT <sub>3</sub>GT<sub>6</sub> allele is also associated with higher systolic BP (beta = 3.3 mmHg, <i>p</i> = 0.014) and diastolic BP (beta = 2.4 mmHg, <i>p</i> = 0.003) in a European-American population. Therefore, we have identified a novel multi-allelic TRS in the 3’UTR of <i>ATP1B1</i> that is associated with higher BP and may mediate its effect by regulating the polyadenylation of the <i>ATP1B1</i> mRNA.</p> </div>", "links"=>[], "tags"=>["polymorphic", "regulates", "polyadenylation"], "article_id"=>812225, "categories"=>["Biological Sciences"], "users"=>["Megana K. Prasad", "Kavita Bhalla", "Zhen Hua Pan", "Jeffrey R. O’Connell", "Alan B. Weder", "Aravinda Chakravarti", "Bin Tian", "Yen-Pei C. Chang"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0076290.s001", "https://dx.doi.org/10.1371/journal.pone.0076290.s002", "https://dx.doi.org/10.1371/journal.pone.0076290.s003", "https://dx.doi.org/10.1371/journal.pone.0076290.s004", "https://dx.doi.org/10.1371/journal.pone.0076290.s005"], "stats"=>{"downloads"=>0, "page_views"=>0, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_Polymorphic_3_UTR_Element_in_ATP1B1_Regulates_Alternative_Polyadenylation_and_Is_Associated_with_Blood_Pressure/812225", "title"=>"A Polymorphic 3’UTR Element in <i>ATP1B1</i> Regulates Alternative Polyadenylation and Is Associated with Blood Pressure", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-10-01 02:12:07"}

PMC Usage Stats | Further Information

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Relative Metric

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