UV‐Induced Changes in Cell Cycle and Gene Expression within Rabbit Lens Epithelial Cells

Sidjanin, D. J.; Grdina, David J.; Woloschak, Gayle E.

doi:10.1111/j.1751-1097.1996.tb02995.x

Cited by 10 publications

(6 citation statements)

References 33 publications

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“…45,46 It has also been reported that histone transcripts accumulate with exposure of lens epithelial cells to ultraviolet C radiation. 47 The possible association of αAand αB-crystallin with histones is also consistent with their involvement in cell growth. 9,11 We further demonstrated that the absence of α-crystallins increases the amount of vimentin cross-linked with tubulin.…”

Section: ■ Discussionmentioning

confidence: 57%

“…Histone H2B undergoes phosphorylation in response to apoptotic signals in mammalian cells, which may facilitate access of damaged DNA to repair mechanisms (44, 45). It has also been reported that histone transcripts accumulate with exposure of lens epithelial cells to ultraviolet C radiation (46). The possible association of αA-and αB-crystallin with histones is also consistent with their involvement in cell growth (9, 11).…”

Section: Discussionmentioning

confidence: 99%

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Comparative Proteomic Analysis Identifies Age-Dependent Increases in the Abundance of Specific Proteins after Deletion of the Small Heat Shock Proteins αA- and αB-Crystallin

Andley¹,

Malone²,

Hamilton

et al. 2013

Biochemistry

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Mice with deletion of genes for small heat shock proteins αA- and αB-crystallin (αA/αB−/−) develop cataracts. We used proteomic analysis to identify lens proteins that change in abundance after deletion of these α-crystallin genes. Wild-type (WT) and αA/αB−/− knockout (DKO) mice were compared using two-dimensional difference gel electrophoresis and mass spectrometric analysis, and protein identifications were validated by Mascot proteomic software. The abundance of histones H2A, H4, and H2B fragment, and a low molecular weight β1-catenin increased 2- to 3-fold in postnatal day 2 lenses of DKO lenses compared with WT lenses. Additional major increases were observed in abundance of βB2-crystallin and vimentin in 30-day-old lenses of DKO animals compared with WT animals. Lenses of DKO mice were comprised of 9 protein spots containing βB2-crystallin at 10- to 40-fold higher abundance and 3 protein spots containing vimentin at ≥ 2-fold higher abundance than in WT lenses. Gel permeation chromatography identified a unique 328 kDa protein in DKO lenses, containing β-crystallin, demonstrating aggregation of β-crystallin in the absence of α-crystallins. Together, these changes provide biochemical evidence for possible functions of specific cell adhesion proteins, cytoskeletal proteins, and crystallins in lens opacities caused by the absence of the major chaperones, αA- and αB-crystallins.

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Section: ■ Discussionmentioning

confidence: 57%

Section: Discussionmentioning

confidence: 99%

Comparative Proteomic Analysis Identifies Age-Dependent Increases in the Abundance of Specific Proteins after Deletion of the Small Heat Shock Proteins αA- and αB-Crystallin

Andley¹,

Malone²,

Hamilton

et al. 2013

Biochemistry

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“…␣A-crystallin has been shown to undergo phosphorylation in vivo and act as an autokinase (52). It is also noteworthy that histone transcripts accumulate in lens epithelial cells after UVC irradiation (53). The phosphorylation of heat-shock protein 60 regulates its attachment to histone H2B in the plasma membrane (54), and a nucleosome assembly protein has recently been shown to function as a histone chaperone (55).…”

Section: Figmentioning

confidence: 99%

The R116C Mutation in αA-crystallin Diminishes Its Protective Ability against Stress-induced Lens Epithelial Cell Apoptosis

Andley¹,

Patel²,

Xi³

2002

Journal of Biological Chemistry

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alphaA-crystallin is a small heat-shock protein expressed preferentially in the lens and is detected during the early stages of lens development. Recent work indicates that the expression of alphaA-crystallin enhances lens epithelial cell growth and resistance to stress conditions. Mutation of the arginine 116 residue to cysteine (R116C) in alphaA-crystallin has been associated with congenital cataracts in humans. However, the physiological consequences of this mutation have not been analyzed in lens epithelial cells. In the present study, we expressed wild type or R116C alphaA-crystallin in the human lens epithelial cell line HLE B-3. Immunofluorescence and confocal microscopy indicated that both wild type and R116C alphaA-crystallin were distributed mainly in the cytoplasm of lens epithelial cells. Size-exclusion chromatography indicated that the size of the alphaA-crystallin aggregate in lens epithelial cells increased from 500 to 600 kDa for the wild type protein to >2 MDa in the R116C mutant. When cells were exposed to physiological levels of UVA radiation, wild type alphaA-crystallin protected cells from apoptotic death as shown by annexin labeling and flow cytometric analysis, whereas the R116C mutant had a 4- to 10-fold lower protective ability. UVA-irradiated cells expressing the wild type protein had very low TUNEL (terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling) staining, whereas cells expressing R116C mutant had a high level of TUNEL staining. F-actin was protected in UVA-treated cells expressing the wild type alphaA-crystallin but was either clumped around the apoptotic cells or was absent in apoptotic cells in cultures expressing the R116C mutant. Structural changes caused by the R116C mutation could be responsible for the reduced ability of the mutant to protect cells from stress. Our study shows that comparing the stress-induced apoptotic cell death is an effective way to compare the protective abilities of wild type and mutant alphaA-crystallin. We propose that the diminished protective ability of the R116C mutant in lens epithelial cells may contribute to the pathogenesis of cataract.

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“…Histones play a central role in the packaging of DNA into nucleosomes, comprise the basic building block of chromatin, and are involved in transcriptional regulation [ 11 – 13 ]; however, very little is known about the specific role of histones in the lens [ 14 , 15 ]. The expression of histone transcripts decreases in lens epithelial cells exposed to UV radiation, suggesting a response to stress [ 16 ]. Also, a recent study demonstrated that the specific acetylation of histone H3 is required for normal lens induction in the embryonic mouse eye at stage E9.5 [ 14 ].…”

Section: Introductionmentioning

confidence: 99%

Probing the changes in gene expression due to α-crystallin mutations in mouse models of hereditary human cataract

et al. 2018

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The mammalian eye lens expresses a high concentration of crystallins (α, β and γ-crystallins) to maintain the refractive index essential for lens transparency. Crystallins are long-lived proteins that do not turnover throughout life. The structural destabilization of crystallins by UV exposure, glycation, oxidative stress and mutations in crystallin genes leads to protein aggregation and development of cataracts. Several destabilizing mutations in crystallin genes are linked with human autosomal dominant hereditary cataracts. To investigate the mechanism by which the α-crystallin mutations Cryaa-R49C and Cryab-R120G lead to cataract formation, we determined whether these mutations cause an altered expression of specific transcripts in the lens at an early postnatal age by RNA-seq analysis. Using knock-in mouse models previously generated in our laboratory, in the present work, we identified genes that exhibited altered abundance in the mutant lenses, including decreased transcripts for Clic5, an intracellular water channel in Cryaa-R49C heterozygous mutant lenses, and increased transcripts for Eno1b in Cryab-R120G heterozygous mutant lenses. In addition, RNA-seq analysis revealed increased histones H2B, H2A, and H4 gene expression in Cryaa-R49C mutant lenses, suggesting that the αA-crystallin mutation regulates histone expression via a transcriptional mechanism. Additionally, these studies confirmed the increased expression of histones H2B, H2A, and H4 by proteomic analysis of Cryaa-R49C knock-in and Cryaa;Cryab gene knockout lenses reported previously. Taken together, these findings offer additional insight into the early transcriptional changes caused by Cryaa and Cryab mutations associated with autosomal dominant human cataracts, and indicate that the transcript levels of certain genes are affected by the expression of mutant α-crystallin in vivo.

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UV‐Induced Changes in Cell Cycle and Gene Expression within Rabbit Lens Epithelial Cells

Cited by 10 publications

References 33 publications

Comparative Proteomic Analysis Identifies Age-Dependent Increases in the Abundance of Specific Proteins after Deletion of the Small Heat Shock Proteins αA- and αB-Crystallin

Comparative Proteomic Analysis Identifies Age-Dependent Increases in the Abundance of Specific Proteins after Deletion of the Small Heat Shock Proteins αA- and αB-Crystallin

The R116C Mutation in αA-crystallin Diminishes Its Protective Ability against Stress-induced Lens Epithelial Cell Apoptosis

Probing the changes in gene expression due to α-crystallin mutations in mouse models of hereditary human cataract

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