{"title":"Absolute quantification of BACH1 and BACH2 transcription factors in B and plasma cells reveals their dynamic changes and unique roles.","authors":"Takeshi Kurasawa, Akihiko Muto, Mitsuyo Matsumoto, Kyoko Ochiai, Kazutaka Murayama, Kazuhiko Igarashi","doi":"10.1093/jb/mvae065","DOIUrl":null,"url":null,"abstract":"<p><p>Changes in the absolute protein amounts of transcription factors are important for regulating gene expression during cell differentiation and in responses to changes in the cellular and extracellular environment. However, few studies have focused on the absolute quantification of mammalian transcription factors. In this study, we established an absolute quantification method for the transcription factors BACH1 and BACH2, which are expressed in B cells and regulated by direct heme binding. The method used purified recombinant proteins as controls in Western blotting and was applied to mouse naïve B cells in the spleen, as well as activated B cells and plasma cells. BACH1 was present in naïve B cells at approximately half the levels of BACH2. In activated B cells, BACH1 decreased compared to naïve B cells, while BACH2 increased. In plasma cells, BACH1 increased back to the same extent as in naïve B cells, while BACH2 was not detected. Their target genes Prdm1 and Hmox1 were highly induced in plasma cells. BACH1 was found to undergo degradation with lower concentrations of heme than BACH2. Therefore, BACH1 and BACH2 are similarly abundant in B cells but differ in heme sensitivity, potentially regulating gene expression differently depending on their heme responsiveness.</p>","PeriodicalId":15234,"journal":{"name":"Journal of biochemistry","volume":null,"pages":null},"PeriodicalIF":2.1000,"publicationDate":"2024-09-25","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of biochemistry","FirstCategoryId":"99","ListUrlMain":"https://doi.org/10.1093/jb/mvae065","RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"BIOCHEMISTRY & MOLECULAR BIOLOGY","Score":null,"Total":0}
引用次数: 0
Abstract
Changes in the absolute protein amounts of transcription factors are important for regulating gene expression during cell differentiation and in responses to changes in the cellular and extracellular environment. However, few studies have focused on the absolute quantification of mammalian transcription factors. In this study, we established an absolute quantification method for the transcription factors BACH1 and BACH2, which are expressed in B cells and regulated by direct heme binding. The method used purified recombinant proteins as controls in Western blotting and was applied to mouse naïve B cells in the spleen, as well as activated B cells and plasma cells. BACH1 was present in naïve B cells at approximately half the levels of BACH2. In activated B cells, BACH1 decreased compared to naïve B cells, while BACH2 increased. In plasma cells, BACH1 increased back to the same extent as in naïve B cells, while BACH2 was not detected. Their target genes Prdm1 and Hmox1 were highly induced in plasma cells. BACH1 was found to undergo degradation with lower concentrations of heme than BACH2. Therefore, BACH1 and BACH2 are similarly abundant in B cells but differ in heme sensitivity, potentially regulating gene expression differently depending on their heme responsiveness.
BACH1 和 BACH2 转录因子在 B 细胞和浆细胞中的绝对定量显示了它们的动态变化和独特作用。
转录因子蛋白质绝对量的变化对细胞分化过程中基因表达的调控以及对细胞和细胞外环境变化的反应非常重要。然而,很少有研究关注哺乳动物转录因子的绝对定量。在这项研究中,我们建立了转录因子 BACH1 和 BACH2 的绝对定量方法,它们在 B 细胞中表达,并通过直接血红素结合进行调控。该方法使用纯化的重组蛋白作为 Western 印迹的对照,适用于小鼠脾脏中的幼稚 B 细胞、活化 B 细胞和浆细胞。BACH1 在幼稚 B 细胞中的含量约为 BACH2 的一半。在活化的 B 细胞中,BACH1 比幼稚 B 细胞减少,而 BACH2 增加。在浆细胞中,BACH1 增加的程度与幼稚 B 细胞相同,而 BACH2 则未检测到。它们的靶基因 Prdm1 和 Hmox1 在浆细胞中被高度诱导。与 BACH2 相比,BACH1 在血红素浓度较低时发生降解。因此,BACH1 和 BACH2 在 B 细胞中的含量相似,但对血红素的敏感性不同,它们可能会根据对血红素的敏感性对基因表达进行不同的调控。
期刊介绍:
The Journal of Biochemistry founded in 1922 publishes the results of original research in the fields of Biochemistry, Molecular Biology, Cell, and Biotechnology written in English in the form of Regular Papers or Rapid Communications. A Rapid Communication is not a preliminary note, but it is, though brief, a complete and final publication. The materials described in Rapid Communications should not be included in a later paper. The Journal also publishes short reviews (JB Review) and papers solicited by the Editorial Board.