Cloning and characterization of the Schizosaccharomyces pombe tRNA:pseudouridine synthase Pus1p

Hellmuth, Klaus; Grosjean, Henri; Motorin, Yuri; Deinert, Karina; Hurt, Ed; Simos, George

doi:10.1093/nar/28.23.4604

Cited by 22 publications

(20 citation statements)

References 27 publications

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“…Previously published experiments using Saccharomyces cerevisiae, Schizosaccharomyces pombe, and mouse homologues of human Pus1p have shown their predominantly nuclear localization (4,29,30). Publicly available prediction programs classified Pus1p as a likely mitochondrial enzyme (10), and Pus1p activity was found in a mitochondrial extract that was shown to be free of cytosolic contaminants (31).…”

Section: Resultsmentioning

confidence: 99%

Mitochondrial Myopathy and Sideroblastic Anemia (MLASA)

Patton

Bykhovskaya

Mengesha

et al. 2005

Journal of Biological Chemistry

124

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A missense mutation in the PUS1 gene affecting a highly conserved amino acid has been associated with mitochondrial myopathy and sideroblastic anemia (MLASA), a rare autosomal recessive oxidative phosphorylation disorder. The PUS1 gene encodes the enzyme pseudouridine synthase 1 (Pus1p) that is known to pseudouridylate tRNAs in other species. Total RNA was isolated from lymphoblastoid cell lines established from patients, parents, unaffected siblings, and unrelated controls, and the tRNAs were assayed for the presence of pseudouridine (⌿) at the expected positions. Mitochondrial and cytoplasmic tRNAs from MLASA patients are lacking modification at sites normally modified by Pus1p, whereas tRNAs from controls, unaffected siblings, or parents all have ⌿ at these positions. In addition, there was no Pus1p activity in an extract made from a cell line derived from a patient with MLASA. Immunohistochemical staining of Pus1p in cell lines showed nuclear, cytoplasmic, and mitochondrial distribution of the protein, and there is no difference in staining between patients and unaffected family members. MLASA is thus associated with absent or greatly reduced tRNA pseudouridylation at specific sites, implicating this pathway in its molecular pathogenesis.Following transcription, all stable RNAs are processed, and nucleotides are modified. The most abundant modification is pseudouridine (⌿), formed by the action of pseudouridine synthases. A number of these modification enzymes have been characterized from several species, and one that has been particularly well studied is pseudouridine synthase 1 (Pus1p).

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Section: Resultsmentioning

confidence: 99%

Mitochondrial Myopathy and Sideroblastic Anemia (MLASA)

Patton

Bykhovskaya

Mengesha

et al. 2005

Journal of Biological Chemistry

124

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“…For example, S. cerevisiae and Schizosaccharomyces pombe Pus1p, which catalyzes U to ⌿ at several tRNA locations and at position 44 of U2 snRNA, is distributed throughout the nucleoplasm (Simos et al 1996;Motorin et al 1998;Massenet et al 1999;Hellmuth et al 2000). In contrast, yeast Trm1p-II, encoding m 2 2 G methyltransferase, and Trm4p, encoding tRNA specific m 5 C methyltransferase, are located at the inner nuclear membrane (INM; Li et al 1989;Rose et al 1995;Wu et al 1998;Motorin and Grosjean 1999).…”

Section: Location Location Locationmentioning

confidence: 99%

tRNA transfers to the limelight

Hopper¹,

Phizicky²

2003

Genes Dev.

292

245

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“…Pseudouridine synthase 1 (Pus1), a member of the TruA family, is a multisite and multisubstrate specific enzyme, which catalyzes pseudouridine formation at several positions of mammalian cytosolic and mitochondrial tRNAs, as well as in snRNA from various species (Arluison et al 1998;Motorin et al 1998;Arluison et al 1999;Chen and Patton 1999;Massenet et al 1999;Chen and Patton 2000;Hellmuth et al 2000;Patton and Padgett 2005;BehmAnsmant et al 2006;Sibert et al 2008). These include positions in the anticodon stem of several tRNAs at positions 27 and 28.…”

Section: Introductionmentioning

confidence: 99%

Formation of a stalled early intermediate of pseudouridine synthesis monitored by real-time FRET

Hengesbach

Voigts-Hoffmann²,

Hofmann³

et al. 2010

RNA

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Pseudouridine is the most abundant of more than 100 chemically distinct natural ribonucleotide modifications. Its synthesis consists of an isomerization reaction of a uridine residue in the RNA chain and is catalyzed by pseudouridine synthases. The unusual reaction mechanism has become the object of renewed research effort, frequently involving replacement of the substrate uridines with 5-fluorouracil (f 5 U). f 5 U is known to be a potent inhibitor of pseudouridine synthase activity, but the effect varies among the target pseudouridine synthases. Derivatives of f 5 U have previously been detected, which are thought to be either hydrolysis products of covalent enzyme-RNA adducts, or isomerization intermediates. Here we describe the interaction of pseudouridine synthase 1 (Pus1p) with f 5 U-containing tRNA. The interaction described is specific to Pus1p and position 27 in the tRNA anticodon stem, but the enzyme neither forms a covalent adduct nor stalls at a previously identified reaction intermediate of f

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Cloning and characterization of the Schizosaccharomyces pombe tRNA:pseudouridine synthase Pus1p

Cited by 22 publications

References 27 publications

Mitochondrial Myopathy and Sideroblastic Anemia (MLASA)

Mitochondrial Myopathy and Sideroblastic Anemia (MLASA)

tRNA transfers to the limelight

Formation of a stalled early intermediate of pseudouridine synthesis monitored by real-time FRET

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