Analysis of the promoters and 5′-UTR of mouse Cd59 genes, and of their functional activity in erythrocytes

Abstract

The complement regulatory protein CD59 inhibits formation of the membrane attack complex (MAC), the terminal effector of the complement system. There are two mouse Cd59 genes in mice but only one in humans. In the work reported here we (a) mapped the promoter regions of both mCd59a and mCd59b genes, (b) identified two different promoters for each mCd59 gene, (c) defined a previously unrecognized additional exon 1 in each mCd59 gene, (d) identified that each mCd59 gene expresses two different tissue-specific transcripts that differ in their 5 0 -UTR, and (e) confirmed the presence of mCd59b mRNA in multiple tissues. At the functional level, comparison of the sensitivity of mCd59ab À/À and mCd59a À/À red blood cells to MAC-mediated lysis revealed that mCd59b protects RBC from MAC-mediated lysis, at least in the setting of mCd59a deficiency. Together these findings indicate that the mCd59 genes may have complex and perhaps different regulatory mechanisms in different tissues.

Nonclassic Functions of Human Topoisomerase I: Genome-Wide and Pharmacologic Analyses

“…Two of those genes belong to the group of nucleic acid and chromatin binding factors, HIST1H2BD (coding for histone H2B type 1-D) and NR2F2, which may compensate for the transcriptional defects resulting from Top1 down-regulation. The two other upregulated genes, CD59 and GIP3, both encode factors that protect cells (44,45). We also found the inactivation in the BAX gene in HCT116-siTop1 cells (data not shown).…”

RETRACTED: The mouse complement regulator CD59b is significantly expressed only in testis and plays roles in sperm acrosome activation and motility

“…Aliquots of these RNAs (1 μg each) were reverse transcribed using random hexamers and multiscribe reverse transcriptase according to the manufacturer's instructions (Applied Biosystems, Warrington, UK). Primer pairs specific for either CD59a or CD59b were as described by Qin et al (2006), and a new primer pair common for both CD59a and b and amplifying a sequence in exon 4 comprising 199 bp for CD59a and 162 bp for CD59b, were used in the subsequent amplifying reactions for detection of CD59a and CD59b mRNAs (Table 1). Either 25 or 35 amplification cycles were used, except where attempting simultaneous detection of both mRNAs when 40 cycles were performed.…”

“…To measure the relative number of mRNA copies for CD59b in liver, perfused liver and testis we used the primer pair reported by Qin et al (2006) to specifically amplify CD59b in a quantitative PCR (QPCR). Because these primers were not designed specifically for this assay, they did not meet the optimal annealing temperature requirements.…”

“…Further, we confirmed that CD59a was dominant in protection of erythrocytes from MAC lysis. The findings of these painstaking studies have recently been challenged yet again (Qin et al, 2006). The appropriateness of the techniques used and specificity of probes and primers have all been questioned.…”

The Role of Complement in the Mechanism of Action of Rituximab for B-Cell Lymphoma: Implications for Therapy

“…It spontaneously identifies any potential pathogens and efficiently protects the host from intruding pathogen attack through a wide range of cellular responses [15][16][17][18]. This system is composed of Ͼ30 soluble plasma proteins and membrane proteins that can trigger three distinct protease cascades known as the classical, mannose-binding lectin (MBL), and alternative pathways (Fig.…”