Comparative genomic analysis of the clade B serpin cluster at human chromosome 18q21: amplification within the mouse squamous cell carcinoma antigen gene locus

Askew, David J.; Askew, Yuko S.; Kato, Yukari; Turner, Russell F.; Dewar, Ken; Lehoczky, Jessica A.; Silverman, Gary A.

doi:10.1016/j.ygeno.2004.01.015

Cited by 34 publications

(36 citation statements)

References 26 publications

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“…These correlations suggest that the functional activity of the clade B serpins has been retained throughout mammalian evolution and that knowledge gained about any one of these genes should help define the overall activity of each orthologous pair. Amino acid analysis of the human and mouse RSLs confirms that these orthologous pairs are likely to demonstrate identical inhibitory profiles (18). Of this set of eight orthologous clade B serpins, only human SERPINB11 and mouse Serpinb11 have yet to be characterized.…”

mentioning

confidence: 74%

“…Serpinb11 was identified using the SER-PINB11 cDNA sequence and BLAST programs to query the mouse genomic and EST data bases as reported (18).…”

Section: Methodsmentioning

confidence: 99%

“…Sequences were analyzed using MacVector 6.0 (Accelrys Inc., Princeton, NJ). Serpinb11 was amplified and cloned from a mouse lung cDNA library (Clontech) as described (18,20).…”

Section: Methodsmentioning

confidence: 99%

“…The clade B cluster at 13A3.2 is greatly expanded (n ϭ 15) relative to that of the human locus at 6p25 (17). Except for the SCCA locus, which contains SERPINB3 and -B4 in humans and Serpinb3a, -b, -c, and -d in mice, the remaining eight clade B serpins mapping to human 18q21 and mouse 1D1 show 1:1 orthology and are highly conserved in terms of gene structure, gene order, and amino acid sequence (18). These correlations suggest that the functional activity of the clade B serpins has been retained throughout mammalian evolution and that knowledge gained about any one of these genes should help define the overall activity of each orthologous pair.…”

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confidence: 99%

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SERPINB11 Is a New Noninhibitory Intracellular Serpin

Askew

Çataltepe

Kumar

et al. 2007

Journal of Biological Chemistry

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SERPINB11, the last of 13 human clade B serpins to be described, gave rise to seven different isoforms. One cDNA contained a premature termination codon, two contained splice variants, and four contained full-length open reading frames punctuated by eight single nucleotide polymorphisms (SNPs). The SNPs encoded amino acid variants located within the serpin scaffold but not the reactive site loop (RSL). Although the mouse orthologue, Serpinb11, could inhibit trypsin-like peptidases, SERPINB11 showed no inhibitory activity. To determine whether the human RSL targeted a different class of peptidases or the serpin scaffold was unable to support inhibitory activity, we synthesized chimeric human and mouse proteins, in which the RSLs had been swapped. The human RSL served as a trypsin inhibitor when supported by mouse scaffold sequences. Conversely, the mouse RSL on the human scaffold showed no inhibitory activity. These findings suggested that variant residues in the SERPINB11 scaffold impaired serpin function. SDS-PAGE analysis supported this notion as RSL-cleaved SERPINB11 was unable to undergo the stressed-to-relaxed transition typical of inhibitory type serpins. Mutagenesis studies supported this hypothesis, since the reversion of amino acid sequences in helices D and I to those conserved in other clade B serpins partially restored the ability of SERPINB11 to form covalent complexes with trypsin. Taken together, these findings suggested that SER-PINB11 SNPs encoded amino acids in the scaffold that impaired RSL mobility, and HapMap data showed that the majority of genomes in different human populations harbored these noninhibitory SERPINB11 alleles. Like several other serpin superfamily members, SERPINB11 has lost inhibitory activity and may have evolved a noninhibitory function.Serpins are a superfamily of serine and cysteine peptidase inhibitors that contain ϳ1500 family members and are found in all domains of life (Eukarya, Eubacteria, and Archaea) as well as many Poxviridae (reviewed in Refs. 1-4). Unlike canonical inhibitors, inhibitory serpins employ a unique suicide substrate-like inhibitory mechanism to neutralize their target peptidases. The surface-exposed reactive site loop (RSL) 4 serves as a pseudosubstrate and binds to the active site of the peptidase. Upon cleavage of the RSL, the metastable serpin molecule undergoes a major conformational rearrangement and traps the covalently attached peptidase in a distorted, inactive form (5). A small proportion of serpins are noninhibitory and aid in diverse functions, such as hormone transport (e.g. SERPINA7/ thyroxine binding globulin) and protein folding (e.g. SER-PINH1/HSP47) (6, 7).Serpins are classified into 17 clades based on phylogenetic relationships (8). So far, 36 human serpins from nine clades (A-I) have been identified (2). The majority of serpins are plasma proteins that serve as critical regulators of important physiological processes, such as blood coagulation, fibrinolysis, and inflammation. In contrast, clade B serpins exist predominantly, but...

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mentioning

confidence: 74%

“…Serpinb11 was identified using the SER-PINB11 cDNA sequence and BLAST programs to query the mouse genomic and EST data bases as reported (18).…”

Section: Methodsmentioning

confidence: 99%

“…Sequences were analyzed using MacVector 6.0 (Accelrys Inc., Princeton, NJ). Serpinb11 was amplified and cloned from a mouse lung cDNA library (Clontech) as described (18,20).…”

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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SERPINB11 Is a New Noninhibitory Intracellular Serpin

Askew

Çataltepe

Kumar

et al. 2007

Journal of Biological Chemistry

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“…The syntenic regions of the mouse genome encode 27 genes, with 15 at chromosome 13 (syntenic to human 6p25) 89 and 12 at chromosome 1 (syntenic to human 18q21). 90 Most of this expansion is due to repeated duplications of genes for SERPINB6 and SERPINB9, from single copies in humans to five and seven paralogues, respectively, in the mouse (Figure 4a). Each additional paralogue has evolved a unique RCL implying non-redundant functions (Figure 4b), and as such the Serpinb9 RCL sequences should show similarity to the predicted substrate specificities of the additional granzymes.…”

Section: Are Rodents Useful For Studying Serpin-granzyme Interactions?mentioning

confidence: 99%

Control of granzymes by serpins

2009

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Although proteolysis mediated by granzymes has an important role in the immune response to infection or tumours, unrestrained granzyme activity may damage normal cells. In this review, we discuss the role of serpins within the immune system, as specific regulators of granzymes. The well-characterised human granzyme B-SERPINB9 interaction highlights the cytoprotective function that serpins have in safeguarding lymphocytes from granzymes that may leak from granules. We also discuss some of the pitfalls inherent in using rodent models of granzyme-serpin interactions and the ways in which our understanding of serpins can help resolve some of the current, contentious issues in granzyme biology. Cell Death and Differentiation (2010) 17, 586-595; doi:10.1038/cdd.2009; published online 6 November 2009As described elsewhere in this series, granzymes are key mediators of cytotoxic lymphocyte (CL) function, exhibiting both intracellular and extracellular functions. Thus, it is imperative that their activities be tightly controlled. Too little activity reduces the effectiveness of the immune system, resulting in infection and cancer. Conversely, overactivity can lead to disease caused by tissue destruction and cellular apoptosis. This review focusses on the manner in which granzyme activity is controlled at the posttranslational level by members of the serpin superfamily.The serpin superfamily is an ever-expanding group of structurally related proteins, currently comprising approximately 1000 members across all kingdoms of life. 1,2 Serpins function as intracellular or extracellular regulators of a wide range of physiological processes such as complement activation, blood coagulation and apoptosis. Within the vertebrate immune system, they control proteases of the classical and innate complement systems, and are also potent inhibitors of many leukocyte granule proteases. Serpin StructureStructures of almost 100 serpins from viruses to humans have been determined and all conform to the same basic architecture first described in 1984. The fold comprises nine a-helices, three b-sheets and a variable reactive centre loop (RCL), which is the primary site of interaction with target proteases (Figure 1a). The first structure determined was human SERPINA1 cleaved within the RCL. 3 Surprisingly, it was found that the residues around the cleavage point were separated by almost 70 Å , with the N-terminal portion of the RCL inserted into b-sheet A to form a fully antiparallel sixstranded sheet. It has subsequently been shown that, in the native state, the RCL is solvent exposed, and that its insertion into b-sheet A is a consequence of the inhibitory mechanism. The RCL is flanked by two highly conserved motifs (the proximal and distal hinges) that permit its insertion into b-sheet A. Insertion is also facilitated by the breach and shutter regions that assist the opening of b-sheet A and by the movement of helix F. 4 The RCL is a critical determinant of serpin inhibitory specificity, acting as a pseudosubstrate and cleavage site for the...

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Study on the targeted therapy of oral squamous cell carcinoma with a plasmid expressing PE38KDEL toxin under control of the SERPINB3 promoter

et al. 2020

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Oral squamous cell carcinoma (OSCC) has a poor prognosis and a high risk of recurrence. To improve the efficacy of OSCC therapy, it is of great significance to explore gene therapy for OSCC. The use of specific genes to regulate the targeted expression of suicide genes is a hot topic in gene therapy for cancer. The SERPINB3 gene is highly active in squamous cell carcinoma, but nearly undetectable or present at a low level in normal tissues. This specificity suggests that the SERPINB3 promoter can be used for targeted OSCC therapy. Pseudomonas aeruginosa secretes PE38KDEL, an exotoxin derivative, as a suicide gene used in gene therapy. A SERPINB3 promoter‐mediated PE38KDEL expression vector was created. The SERPINB3 gene expression was tested in different cell lines by RT‐qPCR and Western blotting, and the SERPINB3 promoter activity was detected by luciferase assay. The SERPINB3 promoter was more active in the TCA8113 cell line than in the other cell lines. The target therapeutic potential of the toxin vector pSERPINB3‐PE38KDEL was tested in the SERPINB3‐positive TCA8113 cell line, the SERPINB3‐negative MG63 cell line, and normal L02 cell line. The SERPINB3 gene was expressed at a high level in TCA8113 cells but a low level in MG63 and L02 cells. Transfection of the pSERPINB3‐PE38KDEL plasmid effectively inhibited the proliferation and invasion of TCA8113 cells and induced cell apoptosis, but no significant damage to MG63 and L02 cells was observed. The results of in vitro experiments indicated that the pSERPINB3‐PE38KDEL plasmid could be a promising strategy for targeted OSCC gene therapy.

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Comparative genomic analysis of the clade B serpin cluster at human chromosome 18q21: amplification within the mouse squamous cell carcinoma antigen gene locus

Cited by 34 publications

References 26 publications

SERPINB11 Is a New Noninhibitory Intracellular Serpin

SERPINB11 Is a New Noninhibitory Intracellular Serpin

Control of granzymes by serpins

Study on the targeted therapy of oral squamous cell carcinoma with a plasmid expressing PE38KDEL toxin under control of the SERPINB3 promoter

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