Cloning and characterization of full-length coding sequence (CDS) of the ovine 37/67-kDa laminin receptor (RPSA)

Abstract

The 37-kDa Laminin Receptor Precursor (LRP)/67-kDa Laminin Receptor (LR), also known as ribosomal protein SA (RPSA), had been identified as a putative cell surface receptor for prions. Herein, we isolated the full-length coding sequence (CDS) of the ovine 37/67-kDa LRP/LR gene and submitted it to the GenBank under accession number EF649775. The open reading frame (ORF) of the 37/67-kDa LRP/LR CDS is 885 bp in length, containing six exons encoding a protein of 295 amino acids. The nucleotide sequence presented here is well coincided with the whole ovine genome of the 37/67-kDa LRP/LR previously published. Moreover, we identified four novel single nucleotide polymorphism sites (SNPs) at position 324 in exon 4, positions at 809, 875, and 881 in exon 7, respectively. Further, based on the deduced amino acid sequence alignment of the 37/67-kDa LRP/LR from human, cattle, mice, pig, chicken, and sheep, we also identified three polymorphic amino acid sites (PAAs) at residues 241, 272, and a novel site at residue 270 in the putative indirect prion protein (PrP) interaction region (180-285) on 37/67-kDa LRP/LR. Prediction of protein secondary structure further indicated that PAAs at residues 241, 270 and 272 may cause protein conformation changes as predicted, which may affect on the binding with prion protein. In addition, multiple-tissues RT-PCR results revealed that 37/67-kDa LRP/LR mRNA is expressed in all the 11 selected ovine tissues.

Molecular Characterization of the Full-Length Coding Sequence of the Caprine Laminin Receptor Gene (RPSA)

“…The housekeeping gene b-actin was used for internal normalization, and the specific primers 5 0 -CTGTCCCTGTACGCCTCT-3 0 (forward) and 5 0 -GTGGTGGTGAA GCTGTAG-3 0 (reverse) were used. The PCR reaction procedure was according to Qiao et al (2008). The PCR product of each tissue was analyzed by electrophoresis on 1% agarose gel and then verified by sequencing.…”

“…Polymorphisms associated with resistance to scrapie infection have been identified in sheep RPSA at interesting amino acid positions (241, 272, and 290), which could participate in the species barrier to scrapie (Marcos-Carcavilla et al 2008). The RPSA gene has been isolated in some species, including human, mouse, cattle, chicken, pig, and sheep (Yow et al 1988;Rao et al 1989;Grosso et al 1991;Clausse et al 1996;Knorr et al 2007;Marcos-Carcavilla et al 2008;Qiao et al 2008). In humans, 17 copies of the human 37 kDa laminin receptor precursor are processed pseudogenes arising from retropositional events .…”

Ribosomal protein SA and its pseudogenes in ruminants: an extremely conserved gene family

“…They neither found polymorphisms that cause an amino acid change. The amino acid sequence of a local Chinese breed deposited by Qiao et al (2009) however differs in four amino acids with the sequences mentioned above. These possible polymorphic amino acids cannot be confirmed by any EST of the NCBI databank.…”

RPSA Gene Mutants Associated with Risk of Colorectal Cancer among the Chinese Population

“…EGCG was also reported to have anti-oxidative (Sang et al, 2005), antimutagenic (Wang et al, 1989), anti-inflammatory (Lin and JK Lin, 1997), and anti-carcinogenic activities (Qiao et al, 2009). However, the effects of tea consumption on CRC are inconsistent in previous epidemiologic studies.…”

“…Ribosomal protein SA (RPSA), which is also called 37 kDa laminin receptor precursor/67 kDa laminin receptor, is a protein with 295 amino acids coded by RPSA gene and the amino acid sequence of RPSA shares high homology in mammals (Qiao et al, 2009). RPSA has been notably identified as a cell-surface EGCG receptor capable of mediating the anti-tumor effects of EGCG in vivo (Tachibana et al, 2004;Umeda et al, 2008;Tsukamoto et al, 2012).…”