Carlo Renieri from the University of Camerino, Italy. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.įunding: This huge in-house project on ‘Molecular Characterization of Pigmentation Genes in Merino Sheep’ was supported by the Faculty Research Grant to Dr. Received: AugAccepted: Published: June 15, 2012Ĭopyright: © 2012 Saravanaperumal et al. PLoS ONE 7(6):Įditor: Jack Anthony Gilbert, Argonne National Laboratory, United States of America This work provides a basis for understanding the functional role and regulation of SCF in hair follicle melanogenesis in sheep beyond what was known in mice, humans and other mammals.Ĭitation: Saravanaperumal SA, Pediconi D, Renieri C, La Terza A (2012) Skipping of Exons by Premature Termination of Transcription and Alternative Splicing within Intron-5 of the Sheep SCF Gene: A Novel Splice Variant. Our data refine the structure of SCF gene clarify the presence (+) and/or absence (−) of primary proteolytic-cleavage site specific SCF splice variants. We also demonstrated that the Northern blot analysis at transcript level is mediated via an intron-5 splicing event. This alternative splice (AS) variant was explained by the complete nucleotide sequencing of splice junction covering exon 5-intron (5)-exon 6 (948 bp) with a premature termination codon (PTC) whereby exons 6 to 9/10 are skipped (Cassette Exon, CE 6–9/10).
It contains an open reading frame (ORF) corresponding to a truncated protein of 181 aa ( vs 245 aa) with an unique C-terminus lacking the primary proteolytic segment (28 aa) right after the D 175G site which is necessary to produce ‘soluble’ form of SCF. In contrast, the shorter (835 and/or 725 bp) cDNA was found to be a ‘novel’ mRNA splice variant. Nucleotide sequencing and molecular prediction revealed that the primary 1519 base pair (bp) cDNA encodes a precursor protein of 274 amino acids (aa), commonly known as ‘soluble’ isoform. Full-length cDNA libraries were enriched by the method of rapid amplification of cDNA ends (RACE-PCR). Reverse transcriptase (RT)-PCR and molecular prediction revealed two different cDNA products of SCF. We report for the first time, a novel mRNA splice variant of SCF from the skin of white merino sheep via cloning and sequencing.
Skin expression of SCF stimulates melanocyte migration, proliferation, differentiation, and survival. In the hematopoietic microenvironment (HM), SCF is produced either as a membrane-bound (−) or soluble (+) forms. Stem cell factor (SCF) is a growth factor, essential for haemopoiesis, mast cell development and melanogenesis.
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