title: Ratios of Four STAT3 Splice Variants in Human Eosinophils and Diffuse Large B Cell Lymphoma Cells.
abstract: Signal transducer and activator of transcription 3 (STAT3) is a key mediator of leukocyte differentiation and proliferation. The 3' end of STAT3 transcripts is subject to two alternative splicing events. One results in either full-length STAT3α or in STAT3β, which lacks part of the C-terminal transactivation domain. The other is at a tandem donor (5') splice site and results in the codon for Ser-701 being included (S) or excluded (ΔS). Despite the proximity of Ser-701 to the site of activating phosphorylation at Tyr-705, ΔS/S splicing has barely been studied. Sequencing of cDNA from purified eosinophils revealed the presence of four transcripts (S-α, ΔS-α, S-β, and ΔS-β) rather than the three reported in publically available databases from which ΔS-β is missing. To gain insight into regulation of the two alternative splicing events, we developed a quantitative(q) PCR protocol to compare transcript ratios in eosinophils in which STAT3 is upregulated by cytokines, activated B cell diffuse large B cell Lymphoma (DLBCL) cells in which STAT3 is dysregulated, and in germinal center B cell-like DLBCL cells in which it is not. With the exception of one line of activated B cell DLCBL cells, the four variants were found in roughly the same ratios despite differences in total levels of STAT3 transcripts. S-α was the most abundant, followed by S-β. ΔS-α and ΔS-β together comprised 15.6 ± 4.0 % (mean ± SD, n = 21) of the total. The percentage of STAT3β variants that were ΔS was 1.5-fold greater than of STAT3α variants that were ΔS. Inspection of Illumina's "BodyMap" RNA-Seq database revealed that the ΔS variant accounts for 10-26 % of STAT3 transcripts across 16 human tissues, with less variation than three other genes with the identical tandem donor splice site sequence. Thus, it seems likely that all cells contain the S-α, ΔS-α, S-β, and ΔS-β variants of STAT3.
authors.affiliation: Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI, United States of America.
authors.forename: Keren B
authors.initials: KB
authors.lastname: Turton
authors.affiliation: Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI, United States of America.
authors.forename: Douglas S
authors.initials: DS
authors.lastname: Annis
authors.affiliation: Department of Medicine at University of Wisconsin-Madison, Madison, WI, United States of America.
authors.forename: Lixin
authors.initials: L
authors.lastname: Rui
authors.affiliation: Department of Medicine at University of Wisconsin-Madison, Madison, WI, United States of America.
authors.forename: Stephane
authors.initials: S
authors.lastname: Esnault
authors.affiliation: Department of Biomolecular Chemistry, University of Wisconsin-Madison, Madison, WI, United States of America; Department of Medicine at University of Wisconsin-Madison, Madison, WI, United States of America.
authors.forename: Deane F
authors.initials: DF
authors.lastname: Mosher
mesh_terms: D017398:Alternative Splicing; D000595:Amino Acid Sequence; D001402:B-Lymphocytes / Q000473:pathology; D001483:Base Sequence; D045744:Cell Line, Tumor; D004804:Eosinophils / Q000378:metabolism*; D018858:Germinal Center / Q000473:pathology; D006801:Humans; D016403:Lymphoma, Large B-Cell, Diffuse / Q000235:genetics*; D008969:Molecular Sequence Data; D022821:RNA Splice Sites / Q000235:genetics; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D015203:Reproducibility of Results; D050796:STAT3 Transcription Factor / Q000737:chemistry / Q000235:genetics*; D014158:Transcription, Genetic
chemical_list: D022821:RNA Splice Sites; D012333:RNA, Messenger; D050796:STAT3 Transcription Factor