BTEB2, a Kruppel-like transcription factor, regulates expression of the SMemb/nonmuscle myosin heavy chain B (SMemb/NMHC-B) gene

Noboru Watanabe, Masahiko Kurabayashi, Yukio Shimomura, Keiko Kawai-Kowase, Yo Ichi Hoshino, Ichiro Manabe, Masafumi Watanabe, Masanori Aikawa, Makoto Kuro-O, Toru Suzuki, Yoshio Yazaki, Ryozo Nagai

Research output: Contribution to journalArticle

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Abstract

We have recently characterized the promoter region of the rabbit embryonic smooth muscle myosin heavy chain (SMemb/NMHC-B) gene and identified the 15-bp sequence, designated SE1, located at -105 from the transcriptional start site as an important regulatory element for its transcriptional activity in a smooth muscle cell (SMC) line. In this study, we attempted to isolate cDNA clones encoding for the transcription factors that control the expression of the SMemb gene through binding to this cis-regulatory element. We screened a λgt11 cDNA library prepared from C2/2 cells, a rabbit-derived SMC line, by using a radiolabeled concatenated oligonucleotide containing SE1 as a probe. Sequence analysis revealed that one of the cDNA clones corresponds to the rabbit homologue of basic transcriptional element binding protein-2 (BTEB2), which has previously been identified as one of the Kruppel-like transcription factor. Gel mobility shift assays and antibody supershift analyses with nuclear extracts from C2/2 cells indicate that BTEB2 is a major component of nuclear factor: SE1 complexes. Furthermore, a glutathione S-transferase-BTEB2 fusion protein binds to the SE1 in a sequence-specific manner. In support of the functionality of BTEB2 binding, basal promoter activity and BTEB2-induced transcriptional activation were markedly attenuated by the disruption of the SE1. In adult rabbit tissues, BTEB2 mRNA was most highly expressed in intestine, urinary bladder, and uterus. BTEB2 mRNA levels were downregulated in rabbit aorta during normal development. Moreover, immunohistochemical analysis indicated a marked induction of BTEB2 protein in the neointimal SMC after balloon injury in rat aorta. These results suggest that BTEB2 mediates the transcriptional regulation of the SMemb/NMHC-B gene and possibly plays a role in regulating gene expression during phenotypic modulation of vascular SMC.

Original languageEnglish (US)
Pages (from-to)182-191
Number of pages10
JournalCirculation Research
Volume85
Issue number2
StatePublished - Jul 23 1999

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Kruppel-Like Transcription Factors
Carrier Proteins
Genes
Smooth Muscle Myocytes
Rabbits
Aorta
Complementary DNA
Clone Cells
Transcriptional Regulatory Elements
Smooth Muscle Myosins
Gene Expression
nonmuscle myosin type IIB heavy chain
Cell Line
Messenger RNA
Myosin Heavy Chains
Electrophoretic Mobility Shift Assay
Glutathione Transferase
Gene Library
Vascular Smooth Muscle
Genetic Promoter Regions

Keywords

  • Basic transcriptional element binding protein-2
  • SMemb/NMHC-B
  • Smooth muscle cell

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Watanabe, N., Kurabayashi, M., Shimomura, Y., Kawai-Kowase, K., Hoshino, Y. I., Manabe, I., ... Nagai, R. (1999). BTEB2, a Kruppel-like transcription factor, regulates expression of the SMemb/nonmuscle myosin heavy chain B (SMemb/NMHC-B) gene. Circulation Research, 85(2), 182-191.

BTEB2, a Kruppel-like transcription factor, regulates expression of the SMemb/nonmuscle myosin heavy chain B (SMemb/NMHC-B) gene. / Watanabe, Noboru; Kurabayashi, Masahiko; Shimomura, Yukio; Kawai-Kowase, Keiko; Hoshino, Yo Ichi; Manabe, Ichiro; Watanabe, Masafumi; Aikawa, Masanori; Kuro-O, Makoto; Suzuki, Toru; Yazaki, Yoshio; Nagai, Ryozo.

In: Circulation Research, Vol. 85, No. 2, 23.07.1999, p. 182-191.

Research output: Contribution to journalArticle

Watanabe, N, Kurabayashi, M, Shimomura, Y, Kawai-Kowase, K, Hoshino, YI, Manabe, I, Watanabe, M, Aikawa, M, Kuro-O, M, Suzuki, T, Yazaki, Y & Nagai, R 1999, 'BTEB2, a Kruppel-like transcription factor, regulates expression of the SMemb/nonmuscle myosin heavy chain B (SMemb/NMHC-B) gene', Circulation Research, vol. 85, no. 2, pp. 182-191.
Watanabe N, Kurabayashi M, Shimomura Y, Kawai-Kowase K, Hoshino YI, Manabe I et al. BTEB2, a Kruppel-like transcription factor, regulates expression of the SMemb/nonmuscle myosin heavy chain B (SMemb/NMHC-B) gene. Circulation Research. 1999 Jul 23;85(2):182-191.
Watanabe, Noboru ; Kurabayashi, Masahiko ; Shimomura, Yukio ; Kawai-Kowase, Keiko ; Hoshino, Yo Ichi ; Manabe, Ichiro ; Watanabe, Masafumi ; Aikawa, Masanori ; Kuro-O, Makoto ; Suzuki, Toru ; Yazaki, Yoshio ; Nagai, Ryozo. / BTEB2, a Kruppel-like transcription factor, regulates expression of the SMemb/nonmuscle myosin heavy chain B (SMemb/NMHC-B) gene. In: Circulation Research. 1999 ; Vol. 85, No. 2. pp. 182-191.
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