Molecular cloning, sequence characteristics, and polymorphism analyses of the tyrosinase-related protein 2 / DOPAchrome tautomerase gene of black-boned sheep (Ovis aries)

Genome ◽  
2009 ◽  
Vol 52 (12) ◽  
pp. 1001-1011 ◽  
Author(s):  
Weidong Deng ◽  
Yuwen Tan ◽  
Xinyu Wang ◽  
Dongmei Xi ◽  
Yiduo He ◽  
...  
Keyword(s):  
Rflp Analysis ◽  
Coat Colour ◽  
Tyrosinase Activity ◽  
Ovis Aries ◽  
Nucleotide Polymorphisms ◽  
Related Protein ◽  
Synonymous Mutations ◽  
Dopachrome Tautomerase ◽  
Flanking Regions ◽  
Tyrosinase Related Protein

Tyrosinase-related protein 2 (TYRP2) plays a pivotal role in the biosynthesis of eumelanin. Black-boned sheep have excessive melanin and eumelanin, resulting in dark (black) muscles and organs. This study was designed to investigate the effects of variants of the TYRP2 gene on black traits and coat colour of black-boned sheep. Melanin traits were measured in three populations of sheep (Nanping black-boned, Nanping normal, and Romney Marsh) and compared in this study. From the TYRP2 cDNA, all 8 exons and their flanking regions were amplified and characterized. Fifteen single nucleotide polymorphisms (SNPs) were identified in the exons and their flanking regions. Five exonic polymorphic sites, including two synonymous (c.93T>G and c.1140C>T) and three non-synonymous mutations (c.163C>T (p.R55W), c.605G>A (p.R202H), and c.1141A>G (p.T381A)), were retrieved. PCR-RFLP analysis of c.605G>A showed that the frequencies of allele G in the Nanping black-boned, Nanping normal, and Romney Marsh sheep were 0.632, 0.603, and 0.886, respectively. Sheep with the GG genotype had significantly (P < 0.05) lower tyrosinase activity, alkali-soluble melanin content, and ratio of eumelanin : total melanin than sheep with GA and AA genotypes when measured across all investigated samples but not when samples within each population of sheep were compared. However, there was no association of TYRP2 genotype at a single SNP position with coat colour across populations. Nonetheless, the two breeds with higher overall tyrosinase activity did produce darker and more varied coat colours than the breed with lower tyrosinase activity.

scholarly journals An examination of melanogenic traits and <i>TYRP1</i> polymorphism in Nanping and Romney Marsh sheep breeds

2018 ◽  
Vol 61 (1) ◽  
pp. 131-141
Author(s):  
Gouzhi Li ◽  
Heli Xiong ◽  
Dongmei Xi ◽  
Sameeullah Memon ◽  
Liping Wang ◽  
...  
Keyword(s):  
Coat Color ◽  
Single Strand Conformation Polymorphism ◽  
Tyrosinase Activity ◽  
Ovis Aries ◽  
Synonymous Mutations ◽  
Allele Specific ◽  
Specific Amplification ◽  
Random Groups ◽  
Flanking Regions ◽  
Conformation Polymorphism

Abstract. The effects of mutations of the gene for tyrosinase-related protein 1 (TYRP1) on the black muscles and coat color in Nanping black-boned sheep were investigated. Tyrosinase activity and melanin content in plasma were measured and compared in three random groups of sheep: Nanping black-boned (101 heads), Nanping normal (106 heads) and Romney Marsh sheep (82 heads, Ovis aries). Eight exons and their partial flanking regions of the TYRP1 gene were amplified. Six intronic mutations and six exonic polymorphisms including two non-synonymous mutations [c.203C > T (p.A68V) and c.1202T > C (p.V401A)] were identified. Using a bi-directional polymerase chain reaction allele-specific amplification (bi-PASA) of the mutation c.203C > T it was shown that the frequencies of allele C in the Nanping black-boned, Nanping normal and Romney Marsh sheep were respectively 0.955, 0.967 and 0.744. For the mutation c.1202T > C, the frequencies of allele T in the three populations of sheep were respectively 0.777, 0.745 and 0.793 as measured using the single-strand conformation polymorphism. When the data from sheep of all three populations with the CC genotype of SNP c.203C > T were pooled, it was found that there was significantly higher (P < 0.05) tyrosinase activity, content of alkali-soluble melanin and ratio of eumelanin : total melanin than in the plasma of sheep with the CT and TT genotypes. This was not so within each of the three groups of sheep. No significant effect of the TRYP1 genotype on coat color was found. Further studies will be necessary to determine the cause of the black traits in Nanping black-boned sheep.

scholarly journals Exploring the potential effect and mechanisms of protocatechuic acid on human hair follicle melanocytes

2020 ◽  
Vol 70 (4) ◽  
pp. 539-549
Author(s):  
Bo Li ◽  
Jun Tan ◽  
Bosheng Zou ◽  
Xiaojia Liu ◽  
Yiling Yu
Keyword(s):  
Hair Follicle ◽  
Human Hair ◽  
Protocatechuic Acid ◽  
Treatment Time ◽  
Tyrosinase Activity ◽  
Dependent Manner ◽  
Related Protein ◽  
Human Hair Follicle ◽  
Dose Dependent ◽  
Tyrosinase Related Protein

AbstractThis study aims to evaluate the effect of protocatechuic acid (PCA) on human hair follicle melanocytes (HFM). Normal primary HFM were isolated and cultured till logarithmic period of second passage, then treated with different concentrations of PCA (0.1–200 μmol L−1) to study the cell proliferation, melanin contents, tyrosinase activity and protein and mRNA expression of melanogenic genes (tyrosinase-related protein 1 (TRP-1), tyrosinase-related protein 2 (TRP-2), and microphthalmia-associated transcription factor (MITF)) in the cultured HFM. In addition, we have also measured the contents of superoxide dismutase (SOD) and glutathione (GSH) in PCA treated HFM. Vitamin C was used as a positive control. The result showed that PCA can decrease the synthesis of melanin and the tyrosinase activity with IC50 = 8.9 μmol L−1 and IC50 = 6.4 μmol L−1, respectively, at the treatment time of 24 hours, without inducing any cytotoxicity in HFM cells. In addition, the mRNA transcription and protein expression levels of TRP-1, TRP-2 and MITF significantly decreased with a dose-dependent manner after 24-hour PCA treated in HFM cells. Furthermore, PCA has significantly increased the SOD and GSH activity in a dose-dependent manner for 24-hour PCA treatment. This study suggested that PCA has an inhibitory effect on the production of melanin through down-regulation of the expression of melanogenesis-related protein and the effect of anti-oxidation, which could be useful for the therapy of melanin overproduction or skin whitening.

Phenotypic rescue of mutant brown melanocytes by a retrovirus carrying a wild-type tyrosinase-related protein gene

Development ◽  
1990 ◽  
Vol 110 (2) ◽  
pp. 471-475 ◽  
Author(s):  
D.C. Bennett ◽  
D. Huszar ◽  
P.J. Laipis ◽  
R. Jaenisch ◽  
I.J. Jackson
Keyword(s):  
Coat Colour ◽  
Specific Protein ◽  
Related Protein ◽  
Histone H4 ◽  
Wild Type ◽  
Leukaemia Virus ◽  
Mouse Cdna ◽  
Phenotypic Rescue ◽  
Moloney Murine Leukaemia Virus ◽  
Tyrosinase Related Protein

A mouse cDNA for the developmentally controlled, melanocyte-specific protein, tyrosinase-related protein 1 (TRP-1), was previously cloned and reported to show genetic linkage with the coat-colour locus brown (b) on mouse chromosome 4. The cDNA has been inserted into a retroviral vector derived from Moloney murine leukaemia virus, under the control of the human histone H4 promoter. This vector was used to infect melanocytes of the immortal line melan-b, which are homozygous for the b mutation and which display light brown pigmentation in culture. Infected cultures containing between 0.2 and 2 copies of provirus per cell displayed an altered phenotype: 20–50% of cells now had the black to dark brown colour characteristic of cultured wild-type (Black, B/B) mouse melanocytes. Thus the TRP-1 gene complements the brown mutation. We conclude that TRP-1 is the product of the wild-type b-locus.

scholarly journals Anti-Melanogenic Effects of Hydroxyectoine via MITF Inhibition by JNK, p38, and AKT Pathways in B16F10 Melanoma Cells

2019 ◽  
Vol 14 (6) ◽  
pp. 1934578X1985852 ◽  
Author(s):  
You C. Chung ◽  
Min-Jin Kim ◽  
Eun Y. Kang ◽  
Yun B. Kim ◽  
Bong S. Kim ◽  
...  
Keyword(s):  
Skin Color ◽  
Inhibitory Effect ◽  
Tyrosinase Activity ◽  
Dependent Manner ◽  
Related Protein ◽  
B16f10 Melanoma ◽  
Melanin Production ◽  
B16f10 Cells ◽  
Dose Dependent ◽  
Tyrosinase Related Protein

Melanin plays a role in determining human skin color of a person, and a large amount of melanin makes the skin color look darkened. The proper amount of melanin formation protects our skin from UV radiation, but excessive melanin production causes hyperpigmentation and leads to freckles, melasma, and lentigo. In this study, we investigated the inhibitory effect of hydroxyectoine on melanogenesis and its mechanism in B16F10 cells. Melanin content and cellular tyrosinase activity were determined. The expression of microphthalmia-associated transcription factor (MITF), and the activities of tyrosinase and other melanogenesis-related enzymes, such as tyrosinase-related protein 1 (TRP-1) and tyrosinase-related protein 2, were also examined. Hydroxyectoine treatment significantly inhibited melanin production and intracellular tyrosinase activity in a dose-dependent manner. Western blot analysis showed that hydroxyectoine also reduced the expressions of tyrosinase and TRP-1. In addition, hydroxyectoine significantly reduced the expression of MITF, a major regulator of melanin production, and inhibited the phosphorylation of p38, c-Jun N-terminal kinase, and activated the protein kinase B. The results demonstrated that hydroxyectoine inhibits the expression of MITF through the inhibition or activation of melanin-related signaling pathways and downregulates melanogenesis by inhibiting melanogenic enzyme expression and tyrosinase activity. Hydroxyectoine has potential value in functional cosmetics applications, such as whitening.

Expression of Tyrosinase-related Protein 2/DOPAchrome Tautomerase in the Retinoblastoma

2001 ◽  
Vol 72 (3) ◽  
pp. 225-234 ◽  
Author(s):  
Tetsuo Udono ◽  
Kazuhiro Takahashi ◽  
Ken-ichi Yasumoto ◽  
Miki Yoshizawa ◽  
Kazuhisa Takeda ◽  
...  
Keyword(s):  
Related Protein ◽  
Dopachrome Tautomerase ◽  
Tyrosinase Related Protein

scholarly journals Melanocytes and Pigmentation Are Affected in Dopachrome Tautomerase Knockout Mice

2004 ◽  
Vol 24 (8) ◽  
pp. 3396-3403 ◽  
Author(s):  
Laurence Guyonneau ◽  
Fabien Murisier ◽  
Anita Rossier ◽  
Alexandre Moulin ◽  
Friedrich Beermann
Keyword(s):  
Knockout Mice ◽  
Null Allele ◽  
Coat Color ◽  
Pigment Epithelium ◽  
Retinal Pigment ◽  
Related Protein ◽  
Melanin Content ◽  
Dopachrome Tautomerase ◽  
Exon 1 ◽  
Tyrosinase Related Protein

ABSTRACT The tyrosinase family comprises three members, tyrosinase (Tyr), tyrosinase-related protein 1 (Tyrp1), and dopachrome tautomerase (Dct). Null mutations and deletions at the Tyr and Tyrp1 loci are known and phenotypically affect coat color due to the absence of enzyme or intracellular mislocalization. At the Dct locus, three mutations are known that lead to pigmentation phenotype. However, these mutations are not null mutations, and we therefore set out to generate a null allele at the Dct gene locus by removing exon 1 of the mouse Dct gene. Mice deficient in Dct [Dcttm1(Cre)Bee ] lack Dct mRNA and dopachrome tautomerase protein. They are viable and do not show any abnormalities in Dct-expressing sites such as skin, retinal pigment epithelium, or brain. However, the mice show a diluted coat color phenotype, which is due to reduced melanin content in hair. Primary melanocytes from Dct knockout mice are viable in culture and show a normal distribution of tyrosinase and tyrosinase-related protein 1. In comparison to the knockout, the slaty mutation (Dctslt /Dctslt ) has less melanin and affects growth of primary melanocytes severely. In summary, we have generated a knockout of the Dct gene in mice with effects restricted to pigment production and coat color.

scholarly journals Tara Tannin Regulates Pigmentation by Modulating Melanogenesis Enzymes and Melanosome Transport Proteins Expression

2020 ◽  
Vol 7 (01) ◽  
pp. e34-e44
Author(s):  
Myra O. Villareal ◽  
Thanyanan Chaochaiphat ◽  
Meriem Bejaoui ◽  
Kozo Sato ◽  
Hiroko Isoda
Keyword(s):  
Skin Color ◽  
Pigment Cell ◽  
Underlying Mechanism ◽  
Mapk Signaling ◽  
Transport Proteins ◽  
Related Protein ◽  
Promoting Effect ◽  
Dopachrome Tautomerase ◽  
Tyrosinase Related Protein

AbstractThe skin color is imparted by the pigment melanin produced in the melanosomes of melanocytes, through the catalytic action of melanogenesis enzymes tyrosinase, tyrosinase-related protein 1, and dopachrome tautomerase. Disruptions in the melanogenesis process may result to hypopigmentation, as observed in cutaneous postinflammatory conditions. Here, the bioactivity of tara tannin, specifically on melanogenesis, was evaluated in vitro using human epidermal melanocytes (HEM) and B16F10 murine melanoma cells in order to determine the possibility that it may be used as a treatment against hypopigmentation. The melanin content of tara tannin-treated B16F10 cells and the expression level of melanogenesis enzymes and melanosome transport proteins were determined. To elucidate the underlying mechanism of tara tannin’s effect on melanogenesis, DNA microarray analysis was performed. Tara tannin significantly increased melanogenesis in both murine and human pigment cell models by upregulating melanogenesis-associated enzymes’ (tyrosinase, tyrosinase-related protein 1, and dopachrome tautomerase) protein and mRNA expression levels, as well as the melanosome transport proteins (myosin Va and RAB27A) expression, both attributed to increased microphthalmia-associated transcription factor (MITF) expression. Global gene expression analysis results revealed the modulation of genes (p≤0.05; fold-change ≥2.0 and ≤−2.0) that are under the transcriptional regulation of MITF and genes relevant for MAPK signaling, metabolic pathways, and cell cycle. Tara tannin has a significant effective melanogenesis-promoting effect, making it a potential therapeutic agent against hypopigmentation disorders. This is the first report on the melanogenesis regulatory effect of tara tannin in vitro.

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