In de novo purine biosynthesis (DNPS), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase (EC 2.1.2.3)/inosine monophosphate cyclohydrolase (EC 3.5.4.10) (ATIC) catalyzes the last two reactions of the pathway: conversion of 5-aminoimidazole-4-carboxamide ribonucleotide [aka Z-nucleotide monophosphate (ZMP)] to 5-formamido-4-imidazolecarboxamide ribonucleotide (FAICAR) then to inosine monophosphate (IMP). Mutations in ATIC cause an untreatable and devastating inborn error of metabolism in humans. ZMP is an adenosine monophosphate (AMP) mimetic and a known activator of AMP-activated protein kinase (AMPK). Recently, a HeLa cell line null mutant for ATIC was constructed via CRISPR-Cas9 mutagenesis. This mutant, crATIC, accumulates ZMP during purine starvation. Given that the mutant can accumulate ZMP in the absence of treatment with exogenous compounds, crATIC is likely an important cellular model of DNPS inactivation and ZMP accumulation. In the current study, we characterize the crATIC transcriptome versus the HeLa transcriptome in purine-supplemented and purine-depleted growth conditions. We report and discuss transcriptome changes with particular relevance to Alzheimer's disease and in genes relevant to lipid and fatty acid synthesis, neurodevelopment, embryogenesis, cell cycle maintenance and progression, extracellular matrix, immune function, TGFβ and other cellular processes.

• Klíčová slova
• 5-aminoimidazole-4-carboxamide ribonucleoside, (AICAr), 5-aminoimidazole-4-carboxamide ribonucleotide formyltransferase/inosine monophosphate cyclohydrolase, (ATIC), 5-aminoimidazole-4-carboxamide ribonucleotide, (ZMP), 5-formamido-4-imidazolecarboxamide ribonucleotide, (FAICAR), AICA-ribosiduria, AMP-activated protein kinase, (AMPK), Alzheimer's disease, Development, Purine synthesis, RNA-seq, Tuberous Sclerosis Complex 1 and 2, (TSC1 and TSC2), adenine phosphoribosyltransferase, (APRT), adenosine monophosphate, (AMP), adenosine triphosphate, (ATP), adenylosuccinate lyase, (ADSL), arachidonic acid, (AA), cyclooxygenase, (COX), cytochrome, P450 (CYP), cytosolic phospholipase A2, (cPLA2), de novo purine synthesis, (DNPS), differentially expressed gene, (DEG), false discovery rate, (FDR), fatty acid amide hydrolase, (FAAH), fetal calf macroserum, (FCM), fetal calf serum, (FCS), fragments per kilobase of exon per million reads mapped, (FPKM), gene ontology, (GO), guanosine monophosphate, (GMP), inosine monophosphate, (IMP), interferon, (INF), lipoxygenase, (LOX), mammalian Target of Rapamycin, (mTOR), minus adenine crATIC to minus adenine WT comparison, (MM), phospholipase, (PLA), phosphoribosyl pyrophosphate, (PRPP), phosphoribosylaminoimidazole carboxylase/phosphoribosylaminoimidazole succinocarboxamide synthetase, (PAICS), plus adenine crATIC to plus adenine WT comparison, (PP), xanthine monophosphate, (XMP),
• Publikační typ
• časopisecké články MeSH

• Klíčová slova
• LC3, autolysosome, autophagosome, chaperone-mediated autophagy, flux, lysosome, macroautophagy, phagophore, stress, vacuole,
• MeSH
• autofagie * fyziologie   MeSH
• biotest metody   normy   MeSH
• lidé MeSH
• počítačová simulace MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Research Support, N.I.H., Extramural MeSH
• směrnice MeSH

Contribution of individual adiponectin isoforms to lipolysis regulation remains unknown. We investigated the impact of full-length, trimeric and globular adiponectin isoforms on spontaneous lipolysis in subcutaneous abdominal (SCAAT) and visceral adipose tissues (VAT) of obese and non-obese subjects. Furthermore, we explored the role of AMPK (5'-AMP-activated protein kinase) in adiponectin-dependent lipolysis regulation and expression of adiponectin receptors type 1 and 2 (AdipoR1 and AdipoR2) in SCAAT and VAT. Primary adipocytes isolated from SCAAT and VAT of obese and non-obese women were incubated with 20 µg/ml of: A) full-length adiponectin (physiological mixture of all adiponectin isoforms), B) trimeric adiponectin isoform or C) globular adiponectin isoform. Glycerol released into media was used as a marker of lipolysis. While full-length adiponectin inhibited lipolysis by 22% in non-obese SCAAT, globular isoform inhibited lipolysis by 27% in obese SCAAT. No effect of either isoform was detected in non-obese VAT, however trimeric isoform inhibited lipolysis by 21% in obese VAT (all p<0.05). Trimeric isoform induced Thr172 p-AMPK in differentiated preadipocytes from a non-obese donor, while globular isoform induced Ser79 p-ACC by 32% (p<0.05) and Ser565 p-HSL by 52% (p = 0.08) in differentiated preadipocytes from an obese donor. AdipoR2 expression was 17% and 37% higher than AdipoR1 in SCAAT of obese and non-obese groups and by 23% higher in VAT of obese subjects (all p<0.05). In conclusion, the anti-lipolytic effect of adiponectin isoforms is modified with obesity: while full-length adiponectin exerts anti-lipolytic action in non-obese SCAAT, globular and trimeric isoforms show anti-lipolytic activity in obese SCAAT and VAT, respectively.

• MeSH
• adiponektin krev   chemie   metabolismus   MeSH
• aminoimidazolkarboxamid analogy a deriváty   farmakologie   MeSH
• dospělí MeSH
• exprese genu účinky léků   MeSH
• hypoglykemika farmakologie   MeSH
• kultivované buňky MeSH
• kvantitativní polymerázová řetězová reakce MeSH
• lidé středního věku MeSH
• lidé MeSH
• lipolýza účinky léků   MeSH
• multimerizace proteinu účinky léků   MeSH
• nitrobřišní tuk cytologie   MeSH
• obezita patologie   MeSH
• podkožní tuk cytologie   MeSH
• protein - isoformy krev   chemie   metabolismus   MeSH
• proteinkinasy aktivované AMP metabolismus   MeSH
• receptory adiponektinu genetika   metabolismus   MeSH
• ribonukleotidy farmakologie   MeSH
• tukové buňky cytologie   účinky léků   metabolismus   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adiponektin MeSH
• ADIPOR1 protein, human MeSH Prohlížeč
• ADIPOR2 protein, human MeSH Prohlížeč
• AICA ribonucleotide MeSH Prohlížeč
• aminoimidazolkarboxamid MeSH
• hypoglykemika MeSH
• protein - isoformy MeSH
• proteinkinasy aktivované AMP MeSH
• receptory adiponektinu MeSH
• ribonukleotidy MeSH