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14-3-3 isoforms bind directly exon B of the 5'-UTR of human surfactant protein A2 mRNA.

ABSTRACT: Human surfactant protein (SP) A (SP-A), an innate immunity molecule, is encoded by two genes, SFTPA1 and SFTPA2. The 5'-untranslated splice variant of SP-A2 (ABD), but not SP-A1 (AD), contains exon B (eB). eB is an enhancer for transcription and translation and contains cis-regulatory elements. Specific trans-acting factors, including 14-3-3, bind eB. The 14-3-3 protein family contains seven isoforms that have been found by mass spectrometry in eB electromobility shift assays (Noutsios et al. Am J Physiol Lung Cell Mol Physiol 304: L722-L735, 2013). We used four different approaches to investigate whether 14-3-3 isoforms bind directly to eB. 1) eB RNA pulldown assays showed that 14-3-3 isoforms specifically bind eB. 2) RNA electromobility shift assay complexes were formed using purified 14-3-3 isoforms ?, ?, ?, ?, ?, and ?, but not isoform ?, with wild-type eB RNA. 3 and 4) RNA affinity chromatography assays and surface plasmon resonance analysis showed that 14-3-3 isoforms ?, ?, ?, ?, ?, and ?, but not isoform ?, specifically and directly bind eB. Inhibition of 14-3-3 isoforms ?, ?, ?, and ?/? with shRNAs in NCI-H441 cells resulted in downregulation of SP-A2 levels but did not affect SP-A1 levels. However, inhibition of 14-3-3 isoform ? was correlated with lower levels of SP-A1 and SP-A2. Inhibition of 14-3-3 isoform ?/?, which does not bind eB, had no effect on expression levels of SP-A1 and SP-A2. In conclusion, the 14-3-3 protein family affects differential regulation of SP-A1 and SP-A2 by binding directly to SP-A2 5'-UTR mRNA.

SUBMITTER: Noutsios GT

PROVIDER: S-EPMC4504974 | biostudies-literature | 2015 Jul

REPOSITORIES: biostudies-literature

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14-3-3 isoforms bind directly exon B of the 5'-UTR of human surfactant protein A2 mRNA.

Noutsios Georgios T GT Ghattas Paul P Bennett Stephanie S Floros Joanna J

American journal of physiology. Lung cellular and molecular physiology 20150522 2

Human surfactant protein (SP) A (SP-A), an innate immunity molecule, is encoded by two genes, SFTPA1 and SFTPA2. The 5'-untranslated splice variant of SP-A2 (ABD), but not SP-A1 (AD), contains exon B (eB). eB is an enhancer for transcription and translation and contains cis-regulatory elements. Specific trans-acting factors, including 14-3-3, bind eB. The 14-3-3 protein family contains seven isoforms that have been found by mass spectrometry in eB electromobility shift assays (Noutsios et al. Am ...[more]

PMID: 26001776

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Dataset Information

14-3-3 isoforms bind directly exon B of the 5'-UTR of human surfactant protein A2 mRNA.

Publications

14-3-3 isoforms bind directly exon B of the 5'-UTR of human surfactant protein A2 mRNA.

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