Isolation of the Regulatory Regions and Genomic Organization of the Porcine ??1,3-Galactosyltransferase Gene1

Koike, Chihiro; Friday, Robert P.; Nakashima, Izumi; Luppi, Patrizia; Fung, John J.; Rao, Abdul S.; Starzl, Thomas E.; Trucco, Massimo

doi:10.1097/00007890-200011150-00004

Cited by 18 publications

(24 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Exon 6Rc in rhesus has exon 6, intron 6, and exon 7 (a retained intron). similar to those in the pig gene (12). In both the rhesus and human, as well as the pig, the region comprises a CpG island.…”

Section: Fig 2 a Electrophoresis Of Pcr (A-c) And Northern Blot Hymentioning

confidence: 81%

“…For this purpose, a fragment of 1431 bp of the 5Ј-flanking region of rhesus exon 1 was inserted into the multicloning site of the pGL3-Basic vector (termed pGL3-RA), which was co-transfected with pRL-TK vector for normalization of transfection efficiency into pig aortic endothelial cells using LipofectAMINE (Invitrogen). Relative luciferase activity of the pGL3-RA was measured by luminometer and compared with that of the pig promoter (12).…”

Section: Methodsmentioning

confidence: 99%

“…Further, the full genomic organization of the porcine ␣1,3GT, gene including its upstream regulatory region, was completed (12). A CpG island (i.e.…”

mentioning

confidence: 99%

“…A CpG island (i.e. a C connected by a 3Ј-5Ј phosphodiester bond to a G) characteristic of housekeeping genes was observed around exon 1 of the pig ␣1,3GT gene (12). It is of interest that a CpG island was not present in the mouse gene (13)(14)(15), suggesting that the upstream regulatory regions might be species-specific even though the enzyme function of the ␣1,3GT gene product was identical (15).…”

mentioning

confidence: 99%

See 3 more Smart Citations

Molecular Basis of Evolutionary Loss of the α1,3-Galactosyltransferase Gene in Higher Primates

Koike¹,

Fung²,

Geller³

et al. 2002

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

Galactose-␣1,3-galactose (␣Gal) epitopes, the synthesis of which requires the enzyme product of ␣1,3-galactosyltransferase (␣1,3GT), are sugar chains on the cell surface of most mammalian species. Notable exceptions are higher primates including Old World monkeys, apes, and humans. The ␣Gal-negative species as well as mice with deletion of the ␣1,3GT gene produce abundant anti␣Gal antibodies. The evolutionary loss of ␣Gal epitopes has been attributed to point mutations in the coding region of the gene. Because no transcripts could be found in the higher primate species with Northern blot analysis, a potential alternative explanation has been loss of upstream regulation of the gene. Here, we have demonstrated that the rhesus promoter is functional. More importantly, a variety of full-length transcripts were detected with sensitive PCR-based methods in the tissues of rhesus monkeys, orangutans, and humans. Five crucial mutations were delineated in the coding region of the human and rhesus and three in the orangutan, any one of which could be responsible for inactivation of the ␣1,3GT gene. Two of the mutations were shared by all three higher primates. These findings, which elucidate the molecular basis for the evolutionary loss of ␣Gal expression, may have implications in medical research.Most mammals express the cell surface carbohydrate epitope galactose-␣1,3-galactose (␣Gal), 1 with notable exceptions that include Old World monkeys, apes, and humans (1). With the loss of the ␣Gal epitopes, the synthesis of which is dependent on enzyme product of the ␣1,3-galactosyltransferase (␣1,3GT) gene, higher primates produce anti-␣Gal antibodies (2) that are responsible for the hyperacute rejection of organs transplanted from ␣Gal-positive donors (3).In 1989, Joziasse et al. (4) reported the sequence of fulllength cDNA clone of the bovine ␣1,3GT gene and demonstrated the presence of this gene in the DNA of human cell lines. Using an 804-bp fragment derived from bovine fulllength cDNA as a probe, they also detected mRNA transcripts in bovine and marmoset (New World monkey) but not in human or African green monkey cell lines (4). The absence of ␣1,3GT mRNA has since been widely viewed as a feature of all Old World monkeys, apes, and humans (5, 6). The molecular basis for the inactivation of the ␣1,3GT gene in the ␣Gal-negative species has been attributed to a mutation(s) localized to a partial sequence of exon 9 (4, 5, 7).Stimulated by the current interest in producing transgenic pigs for clinical use as tissue and organ xenograft donors, the cDNA for the ␣1,3GT gene of the ␣Gal-positive pig was isolated (8), and the coding regions were characterized (9 -12). Further, the full genomic organization of the porcine ␣1,3GT, gene including its upstream regulatory region, was completed (12). A CpG island (i.e. a C connected by a 3Ј-5Ј phosphodiester bond to a G) characteristic of housekeeping genes was observed around exon 1 of the pig ␣1,3GT gene (12). It is of interest that a CpG island was not present in the mouse gene (13-15...

show abstract

Section: Fig 2 a Electrophoresis Of Pcr (A-c) And Northern Blot Hymentioning

confidence: 81%

Section: Methodsmentioning

confidence: 99%

“…Further, the full genomic organization of the porcine ␣1,3GT, gene including its upstream regulatory region, was completed (12). A CpG island (i.e.…”

mentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

Molecular Basis of Evolutionary Loss of the α1,3-Galactosyltransferase Gene in Higher Primates

Koike¹,

Fung²,

Geller³

et al. 2002

Journal of Biological Chemistry

Self Cite

View full text Add to dashboard Cite

show abstract

“…As the first step in clarifying whether any differences exist in the regulation of the al,3GT gene in the two species, we already reported the regulatory regions in pig. 8 Here we report the isolation and characterization of the al,3GT gene promoter regions and the total genomic organization in mice.…”

mentioning

confidence: 99%

Comparison of the regulatory regions of the α1,3galactosyltransferase gene between murine and porcine species

Koike

Friday

Fung

et al. 2001

Transplantation Proceedings

View full text Add to dashboard Cite

Allelic variation of the porcine α-1,3-galactosyltransferase 1 (GGTA1) gene

Day

Rocha

2008

J Appl Genet

View full text Add to dashboard Cite

The alpha-1,3-galactosyltransferase 1 enzyme (GGTA1) produces the alpha-Gal epitopes, responsible for pig-to-human hyperacute xenograft rejection. Recently, efforts have been directed at inactivating the porcine GGTA1 gene in order to reduce hyperacute rejection. As very little is known about the genetic variability of this key gene among pig breeds, we investigated the variation in its nucleotide sequence, by amplification of the entire coding region with the use of polymerase chain reaction followed by DNA sequencing. Eight commercial pig populations were analysed and 17 single nucleotide polymorphisms (SNPs) were detected: 11 in intronic regions and 6 in the 3' untranslated region (UTR). No SNPs change the encoded protein; however, 8 of these SNPs may alter the transcriptional regulation and pre-mRNA splicing of GGTA1.

show abstract

Isolation of the Regulatory Regions and Genomic Organization of the Porcine ??1,3-Galactosyltransferase Gene1

Abstract: These observations have important implications for experiments involving genetic manipulation of the alpha1,3GT gene in transgenic animals in terms of promoter utilization, and particularly in genetically engineering cells for the animal cloning technology by nuclear transfer.

Cited by 18 publications

References 23 publications

Molecular Basis of Evolutionary Loss of the α1,3-Galactosyltransferase Gene in Higher Primates

Molecular Basis of Evolutionary Loss of the α1,3-Galactosyltransferase Gene in Higher Primates

Comparison of the regulatory regions of the α1,3galactosyltransferase gene between murine and porcine species

Allelic variation of the porcine α-1,3-galactosyltransferase 1 (GGTA1) gene

Contact Info

Product

Resources

About