Transcriptional regulation of the chicken ASMT gene - a preliminary analysis.

1. Melatonin is an indole hormone that, among its myriad biological functions, regulates circadian and seasonal rhythms in animals. The ASMT gene plays an essential role in melatonin synthesis. However, in chickens, little is known about the regulatory elements governing its transcription.2. The following study identified the transcription start site of the chicken ASMT gene by 5'-RACE. Then, the proximal minimal promoter was identified using a series of 5' truncations of the ASMT promoter (e.g. -3502/+17, -2698/+17, -2003/+17, -1378/+17, and -254/+17). Site-directed mutagenesis, overexpression, and electrophoretic mobility shift assay (EMSA) were applied to show that the transcription factor Oct-1 binds to the promoter region of ASMT.3. The translation start site was located 19 bp upstream from the translational start site. The luciferase reporter assay confirmed that the core promoter of chicken ASMT gene was in the -254/+17 region. Using site-directed mutagenesis, overexpression, and EMSA, Oct-1 bound the promoter of ASMT.4. Overall, Oct1 plays an important role in the transcriptional regulation of chicken ASMT gene.

Genome Identification of the Tea Plant (Camellia sinensis) ASMT Gene Family and Its Expression Analysis under Abiotic Stress.

The tea plant (Camellia sinensis (L.) O. Ktze) is an important cash crop grown worldwide. It is often subjected to environmental stresses that influence the quality and yield of its leaves. Acetylserotonin-O-methyltransferase (ASMT) is a key enzyme in melatonin biosynthesis, and it plays a critical role in plant stress responses. In this paper, a total of 20 ASMT genes were identified in tea plants and classified into three subfamilies based on a phylogenetic clustering analysis. The genes were unevenly distributed on seven chromosomes; two pairs of genes showed fragment duplication. A gene sequence analysis showed that the structures of the ASMT genes in the tea plants were highly conserved and that the gene structures and motif distributions slightly differed among the different subfamily members. A transcriptome analysis showed that most CsASMT genes did not respond to drought and cold stresses, and a qRT-PCR analysis showed that CsASMT08, CsASMT09, CsASMT10, and CsASMT20 significantly responded to drought and low-temperature stresses; in particular, CsASMT08 and CsASMT10 were highly expressed under low-temperature stress and negatively regulated in response to drought stress. A combined analysis revealed that CsASMT08 and CsASMT10 were highly expressed and that their expressions differed before and after treatment, which indicates that they are potential regulators of abiotic stress resistance in the tea plant. Our results can facilitate further studies on the functional properties of CsASMT genes in melatonin synthesis and abiotic stress in the tea plant.

Polymorphisms in the ASMT and ADAMTS1 gene may increase litter size in goats.

Prolificacy of most local goat breeds in China is low. Jining Grey goat is one of the most prolific goat breeds in China, it is an important goat breed for the rural economy. ASMT (acetylserotonin O-methyltransferase) and ADAMTS1 (ADAM metallopeptidase with thrombospondin type 1 motif) are essential for animal reproduction. Single nucleotide polymorphisms (SNPs) of ASMT and ADAMTS1 genes in the highly prolific breed (Jining Grey goats), medium prolific breed (Boer goats and Guizhou White goats) and low prolific breeds (Angora goats, Liaoning Cashmere goats and Inner Mongolia Cashmere goats) were detected by polymerase chain reaction-restriction fragment length polymorphism and sequencing. Two SNPs (g.158122T>C, g.158700G>A) of ASMT gene and two SNPs (g.7979798A>G, g.7979477C>T) of ADAMTS1 gene were identified. For g.158122T>C of ASMT gene, further analysis revealed that genotype TC or CC had 0.66 (p < 0.05) or 0.75 (p < 0.05) kids more than those with genotype TT in Jining Grey goats. No significant difference (p > 0.05) was found in litter size between TC and CC genotypes. The SNP (g.158122T>C) caused a p.Tyr298His change and this SNP mutation resulted in changes in protein binding sites and macromolecule-binding sites. The improvement in reproductive performance may be due to changes in the structure of ASMT protein. For g.7979477C>T of ADAMTS1 gene, Jining Grey does with genotype CT or TT had 0.82 (p < 0.05) or 0.86 (p < 0.05) more kids than those with genotype CC. No significant difference (p > 0.05) was found in litter size between CT or TT genotypes. These results preliminarily indicated that C allele (g.158122T>C) of ASMT gene and T allele (g.7979477C>T) of ADAMTS1 gene are potential molecular markers which could improve litter size of Jining Grey goats and be used in goat breeding.

An ASMT variant associated with bipolar disorder influences sleep and circadian rhythms: a pilot study.

Patients with bipolar disorder (BD) experience persistent circadian rhythm and sleep abnormalities during periods of remission, and biological studies have shown that these patients have abnormal melatonin secretion profiles or reactivity to light. We previously reported the association with BD of a common polymorphism (rs4446909) of the promoter of the acetylserotonin O-methyltransferase (ASMT) gene, encoding one of the two enzymes involved in melatonin biosynthesis. This variant was associated with weaker transcription and lower levels of ASMT activity in lymphoblastoid cell lines. Actigraphy, based on the use of a mobile portable device for the analysis of sleep/wake cycles in natural conditions, may be useful for studies of carriers of the at-risk allele. We studied the association between the ASMT rs4446909 variant and sleep/activity, as assessed with the Pittsburgh Sleep Quality Index (PSQI) and by actigraphy, in 53 subjects (25 patients with BD in remission and 28 healthy controls). The two groups were similar for age, sex ratio, current mood symptoms, body mass index and risk of sleep apnea syndrome. In the total sample, the GG at-risk genotype was associated with longer sleep duration (P = 0.03), greater activity in active periods of sleep (P = 0.015) and greater interday stability (P = 0.003). These associations remained significant when disease status was included in the model. Only the association with interday stability remained significant after correction for multiple testing. This pilot study thus shows that a BD-associated functional variant involved in the melatonin synthesis pathway influences sleep and circadian rhythms in bipolar patients in remission and controls.