Elsevier

Gene

Volume 376, Issue 1, 5 July 2006, Pages 95-101
Gene

The alleles of PECAM-1

https://doi.org/10.1016/j.gene.2006.02.016Get rights and content

Abstract

Previous studies have reported the existence of eleven different single nucleotide polymorphisms (SNPs) within human PECAM-1 mRNA, several of which have recently been associated with disease. Though SNPs in the PECAM-1 gene have been known for some time, the genetic background on which they exist, and their association into distinct allelic isoforms has not yet been established. To identify the major allelic isoforms of PECAM-1, we determined the nucleotide sequence of individual full-length cloned cDNAs derived from anonymous, unrelated volunteer individuals. Initial sequence analysis of 34 alleles from 17 individuals confirmed the presence of two distinct human PECAM-1 alleles (L98S536R643 and V98N536G643) within the human population. Each of these were found, upon more detailed analysis, to be superimposed on a previously unreported a2479g nucleotide polymorphism within the 3′ untranslated region (3′UTR) that occurred on both allelic isoforms — yielding a total of four major alleles. Multiplex Luminex bead analysis of an additional 259 individuals allowed identification of 117 individuals homozygous for either the L98S536 or V98N536 allele, and sequence analysis around the R643G and a2479g polymorphic sites permitted accurate determination of significant differences in the gene frequencies of LSRa, LSRg, VNGa, and VNGg among Caucasian individuals. Identification of these PECAM-1 allelic isoforms should facilitate future detailed examination of PECAM-1-related disease associations, and may help resolve previously disparate results.

Introduction

PECAM-1 (CD31) is a vascular cell adhesion and signaling receptor that is expressed on the surfaces of platelets, leukocytes, and endothelial cells (Newman et al., 1990), and is encoded by a ∼ 70 kb gene near the end of the long arm of chromosome 17 (17q23) (Gumina et al., 1996). PECAM-1 exists in mature form as a 130 kDa Type I transmembrane glycoprotein comprised of a 574 amino acid extracellular domain containing six Ig-like homology domains, a 19 amino acid transmembrane domain, and a cytoplasmic tail of varying length due to alternative splicing (Kirschbaum et al., 1994). Ig-domain 1 mediates homophilic binding (Liao et al., 1997, Newton et al., 1997, Sun et al., 1996), while Ig-domain 6 binds calcium (Jackson et al., 1997a) and has been suggested to participate in cis interactions with integrin αvβ3 within the plane of the plasma membrane (Wong et al., 2000). The cytoplasmic tail of PECAM-1 possesses two Immunoreceptor Tyrosine Inhibitory Motifs (ITIMs) (Newman, 1999) which, upon tyrosine phosphorylation, recruit and activate the protein tyrosine phosphatase, SHP-2 (Jackson et al., 1997b, Masuda et al., 1997, Sagawa et al., 1997). PECAM-1 has been demonstrated to participate in a variety of physiological events, including leukocyte adhesion and migration, angiogenesis, apoptosis, and modulation of Immunoreceptor Tyrosine Activating Motif (ITAM)-mediated cellular activation (for recent reviews, see (Ilan and Madri, 2003, Newman, 1997, Newman and Newman, 2003)).

Like most genes, variations within the nucleotide sequence of the PECAM-1 gene have been reported, with individual polymorphic residues identified within the 5′UTR, the extracellular and cytoplasmic domains, and the 3′UTR (summarized in Table 1). While the effects of these polymorphisms on PECAM-1-mediated adhesion or signaling have not yet been determined, mismatches at PECAM-1 amino acid residues 98, 536, or 643 have often (Balduini et al., 1999, Behar et al., 1996, Cavanagh et al., 2005, Grumet et al., 2001, Maruya et al., 1998), though not universally (Nichols et al., 1996), been associated with an increased incidence of acute graft-versus-host disease (GVHD), and these and other PECAM-1 polymorphisms have been linked with early onset of atherosclerosis (Elrayess et al., 2003), increased risk of cardiovascular disease (Elrayess et al., 2004, Fang et al., 2005, Listi et al., 2004, Sasaoka et al., 2001, Song et al., 2003, Wei et al., 2004), and susceptibility to malarial infection (Kikuchi et al., 2001), though the latter is also controversial (Casals-Pascual et al., 2001).

While the frequencies of individual PECAM-1 polymorphisms have been determined in a limited number of population studies, these polymorphisms have not, to date, been linked into distinct PECAM-1 alleles, hampering efforts to more definitively establish PECAM-1-related disease associations. The purpose of the present investigation, therefore, was to determine the major alleles bearing each of the most commonly reported SNPs within the PECAM-1 gene. This information should not only permit more precise bio-epidemiological associations to be made amongst different human populations, but also enable biochemical and cell biological studies to be performed to investigate whether functional differences between PECAM-1 allelic isoforms might be causally linked to the reported disease associations of PECAM-1 SNPs.

Section snippets

Preparation of genomic and cDNA from human whole blood

Human whole blood was obtained from anonymous volunteer blood donors. RNA was isolated using a QIAamp® RNA Blood Mini Kit according to manufacturer's instructions (Qiagen, Valencia, CA). cDNA was then prepared from human RNA using the SuperScript™ First-Strand Synthesis for RT-PCR kit (Invitrogen, Carlsbad CA). Following cDNA synthesis, RNase (1 μl) (Invitrogen) was added, tubes incubated at 37 °C for 20 min, and then put on ice or frozen at − 20 °C for later use. Genomic DNA was isolated from

Identification of the primary alleles of human PECAM-1

Single nucleotide polymorphisms within the PECAM-1 gene, including several that result in amino acid substitutions, were originally identified by comparing the sequences of PECAM-1 cDNAs cloned from different laboratories (Newman et al., 1990, Simmons et al., 1990, Stockinger et al., 1990, Zehnder et al., 1992), as well as the single sequence derived thus far from PECAM-1 genomic DNA (Kirschbaum et al., 1994). The frequencies for several individual SNPs have since been determined in several

Acknowledgements

The authors thank Daniel B. Rowe, Ph.D., Division of Biostatistics, Medical College of Wisconsin, for his assistance in statistical analysis. This study was funded by grants HL-40126 and Training Grant HL-07209 from the Heart, Lung, and Blood Institute of the National Institutes of Health grant.

References (42)

  • E. Maruya

    Evidence that CD31, CD49b, and CD62L are immunodominant minor histocompatibility antigens in HLA identical sibling bone marrow transplants

    Blood

    (1998)
  • M. Masuda et al.

    Platelet endothelial cell adhesion molecule-1 is a major SH-PTP2 binding protein in vascular endothelial cells

    FEBS Lett.

    (1997)
  • J.P. Newton et al.

    Residues on both faces of the first immunoglobulin fold contribute to homophilic binding sites on PECAM-1/CD31

    J. Biol. Chem.

    (1997)
  • P. Rousseau et al.

    The 14 bp deletion-insertion polymorphism in the 3′ UT region of the HLA-G gene influences HLA-G mRNA stability

    Hum. Immunol.

    (2003)
  • K. Sagawa et al.

    The protein-tyrosine phosphatase SHP-2 associates with tyrosine-phosphorylated adhesion molecule PECAM-1 (CD31)

    J. Biol. Chem.

    (1997)
  • Q.-H. Sun et al.

    Individually distinct Ig homology domains in PECAM-1 regulate homophilic binding and modulate receptor affinity

    J. Biol. Chem.

    (1996)
  • H. Wei

    Platelet–endothelial cell adhesion molecule-1 gene polymorphism and its soluble level are associated with severe coronary artery stenosis in Chinese Singaporean

    Clin. Biochem.

    (2004)
  • L. Windsor

    Alleles of the IL12B 3′UTR associate with late onset of type 1 diabetes

    Hum. Immunol.

    (2004)
  • J.L. Zehnder et al.

    The cell adhesion molecule CD31 is phosphorylated after cell activation. Down-regulation of CD31 in activated T lymphocytes

    J. Biol. Chem.

    (1992)
  • C.L. Balduini

    Incompatibility for CD31 and human platelet antigens and acute graft-versus-host disease after bone marrow transplantation

    Br. J. Haematol.

    (1999)
  • E. Behar

    Polymorphism of adhesion molecule CD31 and its role in acute graft-versus-host disease

    N. Engl. J. Med.

    (1996)
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