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  • 1
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 20 (1981), S. 6427-6431 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    s.l. : American Chemical Society
    Biochemistry 18 (1979), S. 1438-1442 
    ISSN: 1520-4995
    Source: ACS Legacy Archives
    Topics: Biology , Chemistry and Pharmacology
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-1432
    Keywords: Lens ; Squid ; Crystallin ; Glutathione S-transferase ; Parallel evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Our previous studies have shown that the S-crystallins of cephalopod (Ommastrephes sloani pacificus) eye lenses comprise a family of at least ten members which are evolutionarily related to glutathione S-transferase (GST, EC 2.5.1.18). Here we show by cDNA cloning that there are at least 24 different S-crystallins that are 46–99% identical to each other by amino acid sequence in the squid Loligo opalescens. In each species, all but one S-crystallin (SL11 in O. pacificus and Lops4 in L. opalescens) examined has an inserted central peptide of variable length and sequence. cDNA expression studies conducted in Escherichia coli showed that squid GST (which is expressed little in the lens) has very high enzymatic activity using 1-chloro-2, 4-dinitrobenzene (CDNB) as a substrate; by contrast, SL20-1 of O. pacificus and Lops 12 of L. opalescens (which are encoded by abundant lens mRNAs) have no GST activity. Interestingly, SL11 and Lops4 have some enzymatic activity with the CDNB substrate. Site-specific mutations at Y7 or W38, both residues essential for activity of vertebrate GSTs, or insertion of the central peptide present in the inactive SL20-1, reduced the specific activity of squid GST by 30- to 100-fold. These data indicate that the S-crystallins consist of a family of enzymatically inactive proteins (when using CDNB as a substrate) which is considerably larger than previously believed and that GST activity was lost by gradual drift in sequence as well as by insertion of an extra peptide by exon shuffling. The results are also consistent with the idea that SL11 and Lops4 are orthologous crystallins representing the first descendants of the ancestral GST gene in the pathway which gave rise to the extensive S-crystallin family of lens proteins.
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  • 5
    Electronic Resource
    Electronic Resource
    Springer
    Journal of molecular evolution 33 (1991), S. 495-505 
    ISSN: 1432-1432
    Keywords: Promoter ; Repetitive elements ; 5′-Flanking sequence ; Crystallin genes ; Lens
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Approximately 2 kb of 5′-flanking sequences of the lens-specific αA-crystallin genes from human and mouse are presented and compared with similar regions of the chicken gene. A repetitive element was found approximately 1 kb upstream from the coding sequences of the αA-crystallin gene in all three species (Alu in human, B2 in mouse, and CR1 in chicken), suggesting that they may have an important functional or structural role. Despite the ability of αA-crystallin promoters to function across species, dot matrix analyses show only limited similarity among the 600 bp 5′ to the structural genes of these three species. The human 5′-flanking sequence is more similar to that of the mouse and chicken than the mouse and chicken are to each other. Numerous short sequences (8–13 bp) are common to all three genes but are distributed differently in each species. The locations and conservation of these sequence motifs suggest functional roles, possibly ascis-regulatory elements of transcription. One motif is similar to the αA-CRYBP1 binding site implicated earlier in the transcriptional regulation of the mouse αA-crystallin gene, and other motifs correspond to sites previously mapped by methylation interference studies in the mouse αA-crystallin promoter. The modular arrangement of conserved sequence motifs is consistent with evolutionary changes occurring at the level of gene regulation.
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  • 6
    ISSN: 1432-1432
    Keywords: Lens ; Crystallin ; Squid ; Chicken ; Gene ; Regulation ; AP-1 ; Evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Previous experiments have shown that the minimal promoters required for function of the squid SL20-1 and SL11 crystallin genes in transfected rabbit lens epithelial cells contain an overlapping AP-1/antioxidant responsive element (ARE) upstream of the TATA box. This region resembles the PL-1 and PL-2 elements of the chicken βB 1-cry stallin promoter which are essential for promoter function in transfected primary chicken lens epithelial cells. Here we demonstrate by site-directed mutagenesis that the AP-1/ARE sequence is essential for activity of the squid SL20-1 and SL11 promoters in transfected embryonic chicken lens cells and fibroblasts. Promoter activity was higher in transfected lens cells than in fibroblasts. Electrophoretic mobility shift and DNase protection experiments demonstrated the formation of numerous complexes between nuclear proteins of the embryonic chicken lens and the AP-1/ARE sequences of the squid SL20-1 and SL11 crystallin promoters. One of these complexes comigrated and cross-competed with that formed with the PL-1 element of the chicken βB1-crystallin promoter. This complex formed with nuclear extracts from the lens, heart, brain, and skeletal muscle of embryonic chickens and was eliminated by competition with a consensus AP-1 sequence. The nonfunctional mutant AP-1/ ARE sequences did not compete for complex formation. These data raise the intriguing possibility that entirely different, nonhomologous crystallin genes of the chicken and squid have convergently evolved a similar cis-acting regulatory element (AP-1/ARE) for high expression in the lens.
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  • 7
    ISSN: 1432-1432
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Previous transfection experiments have shown that 162 base pairs (bp) of the 5′ flanking sequence of the chicken αA-crystallin gene are required for promoter activity in primary chicken lens epithelial cells (PLE), while only 111 by of the 5′ flanking sequence are needed for activity of the mouse αA-crystallin promoter in transfected chicken PLE cells or in a SV40 T-antigen-transformed transfected mouse lens epithelial cell line (αTN4-1). The effect of site-directed mutations covering positions −111 to −34 of the mouse αA-crystallin promoter fused to the bacterial chloramphenicol acetyltransferase (CAT) gene was compared in transfected chicken PLE cells and mouse αTN4-1 cells; selected mutations were also examined in a nontransformed rabbit lens epithelial cell line (N/N1003A). In general, the same mutations reduced promoter activity in the transfected lens cells from all three species, although differences were noted. The mutations severely affected regions −111/−106 and −69/−40 regions in all the transfected cells examined; by contrast, mutations at positions −105/−99 and −87/−70 had a somewhat greater effect in the chicken PLE than the mouse αTN4-1 cells, while mutations of the −93/−88 sequence reduced expression in the αTN4-1 but not the PLE cells. A partial cDNA with sequence similarity to αA-CRYPB 1 of the mouse has been isolated from a chicken lens library; mouse αA-CRYBP1 is a putative transcription factor which binds to the −66/−55 sequence of the mouse αA-crystallin promoter. Thus, despite differences in the molecular mechanisms of expression of the chicken and mouse αA-crystallin gene, the present results indicate numerous similarities in the behavior of the mouse promoter in the transfected lens cells of chicken, mouse, and rabbit.
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  • 8
    Electronic Resource
    Electronic Resource
    Oxford, UK : Blackwell Publishing Ltd
    Annals of the New York Academy of Sciences 253 (1975), S. 0 
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1749-6632
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Natural Sciences in General
    Notes: We have described a correlation between the ratio of synthesis of the higher to the lower molecular weight polypeptides of δ-crystallin and the intracellular concentrations of Na+ and K+ in the cultured embryonic chick lens. The only structural differences we have found between the larger and smaller δ-crystallin polypeptides are two, acidic, methionine-containing tryptic peptides which are absent from the lower molecular weight polypeptides. We do not yet know whether the alteration in the ratio of synthesis of the polypeptides is regulated at the post-transcriptional or post-translational level. We are continuing to investigate this problem by studies on the cell-free synthesis of δ-crystallin and by analysis of the sequences and organization of δ-crystallin DNA.One implication of these experiments concerns protein synthesis in cataractous lenses, as has been reviewed elsewhere.15 Numerous cataracts are associated with appreciable changes in ion concentrations, particularly with an increase in Na+ and a decrease in K+. Differential reduction in crystallin synthesis has now been correlated with an increase in the Na+/K+ ratio in three types of cataracts, namely, in hereditary cataracts in mice (Nakano mouse),29 in galactose cataracts in rats,30 and in drug-induced experimental cataracts in rats.31 It seem reasonable to propose that changes in ion concentrations will be correlated with alterations in protein synthesis in other cataracts and. possibly, with other diseases which result in electrolyte imbalances within cells.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Journal of comparative physiology 164 (1989), S. 577-587 
    ISSN: 1432-1351
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Summary The ultrastructure and major soluble proteins of the transparent eye lens of two cubomedusan jellyfish,Tripedalia cystophora andCarybdea marsupialis, have been examined. Each species has two complex eyes (one large and one small) on four sensory structures called rhopalia. The lenses consist of closely spaced cells with few organelles. The lens is situated next to the retina, with only an acellular layer separating it from the photoreceptors. SDS-PAGE showed that the large lens ofC. marsupialis has only two crystallin polypeptide bands (with molecular masses of approximately 20000 and 35000 daltons), while that ofT. cystophora has three bands (two with a molecular mass near 20000 daltons and one with a molecular mass near 35000 daltons). Interestingly, the small lens ofT. cystophora appears to be markedly deficient in or lack the lower molecular weight proteins. The crystallins behaved as monomeric proteins by FPLC and showed no immunological reaction with antisera of the major squid crystallin, chickenδ-crystallin or mouseγ-crystallin in western immunoblots. Very weak reactions were found with antimouseα- andβ-crystallin sera. The 35000 dalton crystallin ofT. cystophora was purified and called J1-crystallin. It contained relatively high leucine (13%) and tyrosine (9%) and low methionine (2%). Several tryptic peptides were sequenced. Weak sequence similarities were found withα- andβ-crystallins, which may account for some of the apparent weak immunological crossreactivity with these vertebrate crystallins. A polyclonal antiserum made in rabbits from a synthetic peptide of J1-crystallin reacted strongly with J1-crystallin ofT. cystophora andC. marsupialis in immunoblots; by contrast, no reaction was obtained with the lower molecular weight crystallins from these jellyfish, with the squid crystallin, or with any crystallins from the frog or human lens. Thus, despite the structural similarities between the cubomedusan, squid and vertebrate lenses, their crystallins appear very different.
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