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Molecular identification and expression analysis of filaggrin-2, a member of the S100 fused-type protein family.

Wu Z, Hansmann B, Meyer-Hoffert U, Gläser R, Schröder JM - PLoS ONE (2009)

Bottom Line: We found that FLG2 transcripts are present in skin, thymus, tonsils, stomach, testis and placenta.We provide evidences that like filaggrin, FLG2 is initially expressed by upper granular cells, proteolytically processed and deposited in the stratum granulosum and stratum corneum (SC) layers of normal epidermis.Thus, FLG2 and filaggrin may have overlapping and perhaps synergistic roles in the formation of the epidermal barrier, protecting the skin from environmental insults and the escape of moisture by offering precursors of natural moisturizing factors.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Hospital of Schleswig-Holstein, Kiel, Germany.

ABSTRACT
Genes of the S100 fused-type protein (SFTP) family are clustered within the epidermal differentiation complex and encode essential components that maintain epithelial homeostasis and barrier functions. Recent genetic studies have shown that mutations within the gene encoding the SFTP filaggrin cause ichthyosis vulgaris and are major predisposing factors for atopic dermatitis. As a vital component of healthy skin, filaggrin is also a precursor of natural moisturizing factors. Here we present the discovery of a member of this family, designated as filaggrin-2 (FLG2) that is expressed in human skin. The FLG2 gene encodes a histidine- and glutamine-rich protein of approximately 248 kDa, which shares common structural features with other SFTP members, in particular filaggrin. We found that FLG2 transcripts are present in skin, thymus, tonsils, stomach, testis and placenta. In cultured primary keratinocytes, FLG2 mRNA expression displayed almost the same kinetics as that of filaggrin following Ca(2+) stimulation, suggesting an important role in molecular regulation of epidermal terminal differentiation. We provide evidences that like filaggrin, FLG2 is initially expressed by upper granular cells, proteolytically processed and deposited in the stratum granulosum and stratum corneum (SC) layers of normal epidermis. Thus, FLG2 and filaggrin may have overlapping and perhaps synergistic roles in the formation of the epidermal barrier, protecting the skin from environmental insults and the escape of moisture by offering precursors of natural moisturizing factors.

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Expression analyses of FLG2 transcripts.(A) Expression profile of FLG2 and FLG mRNA. Fragments were obtained after RT-PCR amplification on multiple human tissue cDNAs with primers specific to human FLG2, FLG and GAPDH. Lanes are labelled according to the template tissue. M, DNA molecular weight marker. (B) FLG2 mRNA is expressed at various localizations of human skin. The mRNA expression level of FLG2 was measured by quantitative real-time PCR in human skin samples. Data were obtained from three independent experiments with different samples of skin and are indicated as absolute mRNA copy number per 10 ng of total RNA and as the mean+/−SD. Feet, the sole of the foot. (C–D) Comparative analyses of FLG2 (C) and FLG (D) expression in cultured primary keratinocytes. Quantitative real-time PCR was conducted on total RNA samples collected from keratinocytes treated with 1.0 mM CaCl2 for the indicated times. Bar graphs represent the relative mRNA expression of either FLG2 or FLG against GAPDH. Data are obtained from three independent trials with different sources of keratinocytes and are indicated as the mean+/−SD.
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pone-0005227-g004: Expression analyses of FLG2 transcripts.(A) Expression profile of FLG2 and FLG mRNA. Fragments were obtained after RT-PCR amplification on multiple human tissue cDNAs with primers specific to human FLG2, FLG and GAPDH. Lanes are labelled according to the template tissue. M, DNA molecular weight marker. (B) FLG2 mRNA is expressed at various localizations of human skin. The mRNA expression level of FLG2 was measured by quantitative real-time PCR in human skin samples. Data were obtained from three independent experiments with different samples of skin and are indicated as absolute mRNA copy number per 10 ng of total RNA and as the mean+/−SD. Feet, the sole of the foot. (C–D) Comparative analyses of FLG2 (C) and FLG (D) expression in cultured primary keratinocytes. Quantitative real-time PCR was conducted on total RNA samples collected from keratinocytes treated with 1.0 mM CaCl2 for the indicated times. Bar graphs represent the relative mRNA expression of either FLG2 or FLG against GAPDH. Data are obtained from three independent trials with different sources of keratinocytes and are indicated as the mean+/−SD.

Mentions: To determine the expression profile of FLG2, we first compared its expression with that of the FLG gene by using reverse transcriptase PCR (RT-PCR) analyses. As shown in Fig. 4A, both transcripts were detected in skin, thymus, stomach, tonsils, testis and placenta, but not in heart, brain, liver, lung, bone marrow, small intestine, spleen, prostate, colon, or adrenal gland (data not shown). Whereas FLG transcripts were detected in kidney, pancreas, mammary gland, bladder, thyroid, salivary gland and trachea, FLG2 transcripts were not (Fig. 4A). To further analyze FLG2 gene expression in human healthy skin, various tissue specimens from different body sites were investigated by quantitative real-time PCR. FLG2 was expressed in all normal skin samples tested (Fig. 4B). In addition, FLG2 mRNA level did not show significant differences among different localizations investigated and its absolute copy number was about 6–30 copies per 10 ng of total RNA.


Molecular identification and expression analysis of filaggrin-2, a member of the S100 fused-type protein family.

Wu Z, Hansmann B, Meyer-Hoffert U, Gläser R, Schröder JM - PLoS ONE (2009)

Expression analyses of FLG2 transcripts.(A) Expression profile of FLG2 and FLG mRNA. Fragments were obtained after RT-PCR amplification on multiple human tissue cDNAs with primers specific to human FLG2, FLG and GAPDH. Lanes are labelled according to the template tissue. M, DNA molecular weight marker. (B) FLG2 mRNA is expressed at various localizations of human skin. The mRNA expression level of FLG2 was measured by quantitative real-time PCR in human skin samples. Data were obtained from three independent experiments with different samples of skin and are indicated as absolute mRNA copy number per 10 ng of total RNA and as the mean+/−SD. Feet, the sole of the foot. (C–D) Comparative analyses of FLG2 (C) and FLG (D) expression in cultured primary keratinocytes. Quantitative real-time PCR was conducted on total RNA samples collected from keratinocytes treated with 1.0 mM CaCl2 for the indicated times. Bar graphs represent the relative mRNA expression of either FLG2 or FLG against GAPDH. Data are obtained from three independent trials with different sources of keratinocytes and are indicated as the mean+/−SD.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2668185&req=5

pone-0005227-g004: Expression analyses of FLG2 transcripts.(A) Expression profile of FLG2 and FLG mRNA. Fragments were obtained after RT-PCR amplification on multiple human tissue cDNAs with primers specific to human FLG2, FLG and GAPDH. Lanes are labelled according to the template tissue. M, DNA molecular weight marker. (B) FLG2 mRNA is expressed at various localizations of human skin. The mRNA expression level of FLG2 was measured by quantitative real-time PCR in human skin samples. Data were obtained from three independent experiments with different samples of skin and are indicated as absolute mRNA copy number per 10 ng of total RNA and as the mean+/−SD. Feet, the sole of the foot. (C–D) Comparative analyses of FLG2 (C) and FLG (D) expression in cultured primary keratinocytes. Quantitative real-time PCR was conducted on total RNA samples collected from keratinocytes treated with 1.0 mM CaCl2 for the indicated times. Bar graphs represent the relative mRNA expression of either FLG2 or FLG against GAPDH. Data are obtained from three independent trials with different sources of keratinocytes and are indicated as the mean+/−SD.
Mentions: To determine the expression profile of FLG2, we first compared its expression with that of the FLG gene by using reverse transcriptase PCR (RT-PCR) analyses. As shown in Fig. 4A, both transcripts were detected in skin, thymus, stomach, tonsils, testis and placenta, but not in heart, brain, liver, lung, bone marrow, small intestine, spleen, prostate, colon, or adrenal gland (data not shown). Whereas FLG transcripts were detected in kidney, pancreas, mammary gland, bladder, thyroid, salivary gland and trachea, FLG2 transcripts were not (Fig. 4A). To further analyze FLG2 gene expression in human healthy skin, various tissue specimens from different body sites were investigated by quantitative real-time PCR. FLG2 was expressed in all normal skin samples tested (Fig. 4B). In addition, FLG2 mRNA level did not show significant differences among different localizations investigated and its absolute copy number was about 6–30 copies per 10 ng of total RNA.

Bottom Line: We found that FLG2 transcripts are present in skin, thymus, tonsils, stomach, testis and placenta.We provide evidences that like filaggrin, FLG2 is initially expressed by upper granular cells, proteolytically processed and deposited in the stratum granulosum and stratum corneum (SC) layers of normal epidermis.Thus, FLG2 and filaggrin may have overlapping and perhaps synergistic roles in the formation of the epidermal barrier, protecting the skin from environmental insults and the escape of moisture by offering precursors of natural moisturizing factors.

View Article: PubMed Central - PubMed

Affiliation: Department of Dermatology, University Hospital of Schleswig-Holstein, Kiel, Germany.

ABSTRACT
Genes of the S100 fused-type protein (SFTP) family are clustered within the epidermal differentiation complex and encode essential components that maintain epithelial homeostasis and barrier functions. Recent genetic studies have shown that mutations within the gene encoding the SFTP filaggrin cause ichthyosis vulgaris and are major predisposing factors for atopic dermatitis. As a vital component of healthy skin, filaggrin is also a precursor of natural moisturizing factors. Here we present the discovery of a member of this family, designated as filaggrin-2 (FLG2) that is expressed in human skin. The FLG2 gene encodes a histidine- and glutamine-rich protein of approximately 248 kDa, which shares common structural features with other SFTP members, in particular filaggrin. We found that FLG2 transcripts are present in skin, thymus, tonsils, stomach, testis and placenta. In cultured primary keratinocytes, FLG2 mRNA expression displayed almost the same kinetics as that of filaggrin following Ca(2+) stimulation, suggesting an important role in molecular regulation of epidermal terminal differentiation. We provide evidences that like filaggrin, FLG2 is initially expressed by upper granular cells, proteolytically processed and deposited in the stratum granulosum and stratum corneum (SC) layers of normal epidermis. Thus, FLG2 and filaggrin may have overlapping and perhaps synergistic roles in the formation of the epidermal barrier, protecting the skin from environmental insults and the escape of moisture by offering precursors of natural moisturizing factors.

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Related in: MedlinePlus