Deletions within a CA-repeat-rich region of intron 4 of the human SP-B gene affect mRNA splicing

Zhenwu Lin, Neal J. Thomas, Yunhua Wang, Xiaoxuan Guo, Carola Seifart, Hasan Shakoor, Joanna Floros

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Length variants within a CA-repeat-rich region of intron 4 of the human SP-B (pulmonary surfactantprotein-B) gene are associated with several lung diseases. The hypothesis that SP-B intron 4 affects mRNA splicing was studied. SP-B minigenes containing exons 1-6 with a normal-sized intron 4 (pBi4normal) or intron 4 containing deletions (pBi4del) of 193, 211, 264 or 340 bp were expressed in CHO (Chinese hamster ovary) cells by transient transfection. Two forms of SP-B transcripts, normal and incompletely spliced, were detected. With pBi4normal, normal-sized SP-B mRNA was the predominant form and a very low amount of incompletely spliced mRNA was present, whereas with the pBi4del variants the amount of normal SP-B mRNAs was lower and the amount of incompletely spliced mRNA was relatively high. Reverse transcription-PCR results and sequencing data indicated that the incompletely spliced SP-B RNA contained intron 4 sequence, and this incompletely spliced RNA was also observed in normal lung. Lung cancer tissues with intron 4 deletions exhibited a larger amount of abnormally spliced RNAs compared with normal lung tissue or cancerous tissue with normal-sized intron 4. The results indicate that intron 4 length variants affect SP-B mRNA splicing, and that this may contribute to lung disease.

Original languageEnglish (US)
Pages (from-to)403-412
Number of pages10
JournalBiochemical Journal
Volume389
Issue number2
DOIs
StatePublished - Jul 15 2005

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Introns
Genes
Lung
Messenger RNA
Pulmonary diseases
RNA
Tissue
Lung Diseases
Transcription
Cricetulus
Exons
Reverse Transcription
Transfection
Ovary
Lung Neoplasms
Cells
Polymerase Chain Reaction

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Lin, Zhenwu ; Thomas, Neal J. ; Wang, Yunhua ; Guo, Xiaoxuan ; Seifart, Carola ; Shakoor, Hasan ; Floros, Joanna. / Deletions within a CA-repeat-rich region of intron 4 of the human SP-B gene affect mRNA splicing. In: Biochemical Journal. 2005 ; Vol. 389, No. 2. pp. 403-412.
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abstract = "Length variants within a CA-repeat-rich region of intron 4 of the human SP-B (pulmonary surfactantprotein-B) gene are associated with several lung diseases. The hypothesis that SP-B intron 4 affects mRNA splicing was studied. SP-B minigenes containing exons 1-6 with a normal-sized intron 4 (pBi4normal) or intron 4 containing deletions (pBi4del) of 193, 211, 264 or 340 bp were expressed in CHO (Chinese hamster ovary) cells by transient transfection. Two forms of SP-B transcripts, normal and incompletely spliced, were detected. With pBi4normal, normal-sized SP-B mRNA was the predominant form and a very low amount of incompletely spliced mRNA was present, whereas with the pBi4del variants the amount of normal SP-B mRNAs was lower and the amount of incompletely spliced mRNA was relatively high. Reverse transcription-PCR results and sequencing data indicated that the incompletely spliced SP-B RNA contained intron 4 sequence, and this incompletely spliced RNA was also observed in normal lung. Lung cancer tissues with intron 4 deletions exhibited a larger amount of abnormally spliced RNAs compared with normal lung tissue or cancerous tissue with normal-sized intron 4. The results indicate that intron 4 length variants affect SP-B mRNA splicing, and that this may contribute to lung disease.",
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Deletions within a CA-repeat-rich region of intron 4 of the human SP-B gene affect mRNA splicing. / Lin, Zhenwu; Thomas, Neal J.; Wang, Yunhua; Guo, Xiaoxuan; Seifart, Carola; Shakoor, Hasan; Floros, Joanna.

In: Biochemical Journal, Vol. 389, No. 2, 15.07.2005, p. 403-412.

Research output: Contribution to journalArticle

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