RESUMO
In most organisms, transition metal ions are necessary cofactors of ribonucleotide reductase (RNR), the enzyme responsible for biosynthesis of the 2'-deoxynucleotide building blocks of DNA. The metal ion generates an oxidant for an active site cysteine (Cys), yielding a thiyl radical that is necessary for initiation of catalysis in all RNRs. Class I enzymes, widespread in eukaryotes and aerobic microbes, share a common requirement for dioxygen in assembly of the active Cys oxidant and a unique quaternary structure, in which the metallo- or radical-cofactor is found in a separate subunit, ß, from the catalytic α subunit. The first class I RNRs, the class Ia enzymes, discovered and characterized more than 30 years ago, were found to use a diiron(III)-tyrosyl-radical Cys oxidant. Although class Ia RNRs have historically served as the model for understanding enzyme mechanism and function, more recently, remarkably diverse bioinorganic and radical cofactors have been discovered in class I RNRs from pathogenic microbes. These enzymes use alternative transition metal ions, such as manganese, or posttranslationally installed tyrosyl radicals for initiation of ribonucleotide reduction. Here we summarize the recent progress in discovery and characterization of novel class I RNR radical-initiating cofactors, their mechanisms of assembly, and how they might function in the context of the active class I holoenzyme complex.
Assuntos
Coenzimas , Metais , Ribonucleotídeo Redutases , Animais , Catálise , Domínio Catalítico , Coenzimas/química , Coenzimas/classificação , Coenzimas/metabolismo , Humanos , Metais/química , Metais/metabolismo , Oxirredução , Ribonucleotídeo Redutases/química , Ribonucleotídeo Redutases/classificação , Ribonucleotídeo Redutases/metabolismoRESUMO
The molybdenum cofactor (Moco) represents an ancient metalsulfur cofactor, which participates as catalyst in carbon, nitrogen and sulfur cycles, both on individual and global scale. Given the diversity of biological processes dependent on Moco and their evolutionary age, Moco is traced back to the last universal common ancestor (LUCA), while Moco biosynthetic genes underwent significant changes through evolution and acquired additional functions. In this review, focused on eukaryotic Moco biology, we elucidate the benefits of gene fusions on Moco biosynthesis and beyond. While originally the gene fusions were driven by biosynthetic advantages such as coordinated expression of functionally related proteins and product/substrate channeling, they also served as origin for the development of novel functions. Today, Moco biosynthetic genes are involved in a multitude of cellular processes and loss of the according gene products result in severe disorders, both related to Moco biosynthesis and secondary enzyme functions.
Assuntos
Coenzimas/genética , Eucariotos/genética , Metaloproteínas/genética , Molibdênio/metabolismo , Coenzimas/biossíntese , Coenzimas/classificação , Fusão Gênica/genética , Humanos , Metaloproteínas/biossíntese , Metaloproteínas/classificação , Cofatores de Molibdênio , Pteridinas/classificação , Especificidade por SubstratoAssuntos
Coenzimas/biossíntese , Amina Oxidase (contendo Cobre)/fisiologia , Sequência de Aminoácidos , Animais , Catálise , Códon de Terminação , Coenzimas/química , Coenzimas/classificação , Coenzimas/fisiologia , Cobre , Cristalografia por Raios X , Di-Hidroxifenilalanina/análogos & derivados , Di-Hidroxifenilalanina/biossíntese , Di-Hidroxifenilalanina/química , Dipeptídeos/biossíntese , Endonucleases/fisiologia , Humanos , Indolquinonas/biossíntese , Íons , Dados de Sequência Molecular , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/química , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/genética , Oxirredutases atuantes sobre Doadores de Grupo CH-NH/fisiologia , DNA Polimerase Dirigida por RNA/fisiologia , Triptofano/análogos & derivados , Triptofano/biossínteseRESUMO
The conserved eukaryotic AAA-type ATPase complex, known as p97 or VCP in mammals and Cdc48 in yeast, is involved in a number of cellular pathways, including fusion of homotypic membranes, protein degradation, and activation of membrane-bound transcription factors. Most likely, p97 is directed to this broad spectrum of cellular functions through its binding to specific cofactors. More than 20 different p97 cofactors have been described to date and our understanding of their cellular functions is rapidly expanding. Common to these proteins is their intimate connection with the ubiquitin system. Recently, a small, conserved family of proteins, containing PUB domains, was found to function as p97 adaptors. Intriguingly, their association with p97 is regulated by tyrosine phosphorylation, suggesting that they act as a relay between signalling pathways and p97 functions. Here we give an overview of the currently known PUB-domain proteins and other p97-interacting proteins.