Cloning and transcriptional expression of mouse mannosyltransferase IV/V cDNA, which is involved in the synthesis of lipid-linked oligosaccharides

• Published in: Bioscience, biotechnology, and biochemistry Vol. 78; no. 3; pp. 400 - 409
• Main Authors: Nishimura, Yuuki, Shimono, Nanae, Yoshimoto, Takashi, Kamiguchi, Hiroshi, Nishikawa, Yoshihisa
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01.01.2014; Oxford University Press
• Subjects: alternative donor site selection; alternative splicing; Alternative Splicing - genetics; Animals; Cloning, Molecular; DNA, Complementary - genetics; glycosyltransferase; HeLa Cells; Humans; lipid-linked oligosaccharides; Lipopolysaccharides - biosynthesis; Lipopolysaccharides - genetics; mannosyltransferase; Mannosyltransferases - biosynthesis; Mannosyltransferases - genetics; Mice; Transcription, Genetic; mannosyltransferase; lipid-linked oligosaccharides; alternative splicing; glycosyltransferase; alternative donor site selection
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1080/09168451.2014.890026

We cloned the mouse mannosyltransferase IV/V gene (mALG11) from FM3A cells by a bioinformatic approach. The ORF contained 1476 bp encoding 492 amino acids. The cloned mALG11 complemented the growth defect of the Saccharomyces cerevisiae ALG11Δ mutant. In addition, we detected a variant cDNA by alternate splicing that had an additional four-nucleotide ATGC insertion at base 276 of the ORF. Consequently the variant cDNA encoded a truncated protein with 92 amino acids, lacking the glycosyltransferase group-1 domain. The variant cDNA occurs in many mouse strains according to EST database searches. Moreover, we detected it in FM3A cDNA, but we did not detect any such variants in the human EST database or in HeLa cDNA, although human ALG11 (hALG11) genomic DNA has the same sequence around the intron-exon boundaries as those of mALG11 genomic DNA. Hence, we concluded that there is different transcriptional control mechanism between mALG11 and hALG11. We cloned the mouse mannosyltransferase IV/V gene (mALG11) and its splice-variant from mouse FM3A cells. The latter variant was caused by an alternative donor site selection.