title: Hepatocyte DNA replication in growing liver requires either glutathione or a single allele of txnrd1.
abstract: Ribonucleotide reductase (RNR) activity requires an electron donor, which in bacteria, yeast, and plants is usually either reduced thioredoxin (Trx) or reduced glutaredoxin. Mice lacking glutathione reductase are viable and, although mice lacking thioredoxin reductase 1 (TrxR1) are embryonic-lethal, several studies have shown that mouse cells lacking the txnrd1 gene, encoding TrxR1, can proliferate normally. To better understand the in vivo electron donor requirements for mammalian RNR, we here investigated whether replication of TrxR1-deficient hepatocytes in mouse livers either employed an alternative source of Trx-reducing activity or, instead, solely relied upon the glutathione (GSH) pathway. Neither normal nor genetically TrxR1-deficient livers expressed substantial levels of mRNA splice forms encoding cytosolic variants of TrxR2, and the TrxR1-deficient livers showed severely diminished total TrxR activity, making it unlikely that any alternative TrxR enzyme activities complemented the genetic TrxR1 deficiency. To test whether the GSH pathway was required for replication, GSH levels were depleted by administration of buthionine sulfoximine (BSO) to juvenile mice. In controls not receiving BSO, replicative indexes were similar in hepatocytes having two, one, or no functional alleles of txnrd1. After BSO treatment, hepatocytes containing either two or one copies of this gene were also normal. However, hepatocytes completely lacking a functional txnrd1 gene exhibited severely reduced replicative indexes after GSH depletion. We conclude that hepatocyte proliferation in vivo requires either GSH or at least one functional allele of txnrd1, demonstrating that either the GSH- or the TrxR1-dependent redox pathway can independently support hepatocyte proliferation during liver growth.
authors.affiliation: Department of Immunology and Infectious Disease, Montana State University, Bozeman, MT 59718, USA.
authors.forename: Justin R
authors.initials: JR
authors.lastname: Prigge
authors.forename: Sofi
authors.initials: S
authors.lastname: Eriksson
authors.forename: Sonya V
authors.initials: SV
authors.lastname: Iverson
authors.forename: Tesia A
authors.initials: TA
authors.lastname: Meade
authors.forename: Mario R
authors.initials: MR
authors.lastname: Capecchi
authors.forename: Elias S J
authors.initials: ES
authors.lastname: Arnér
authors.forename: Edward E
authors.initials: EE
authors.lastname: Schmidt
mesh_terms: D000818:Animals; D001483:Base Sequence; D049109:Cell Proliferation; D004261:DNA Replication; D005091:Exons; D005978:Glutathione / Q000378:metabolism / Q000502:physiology*; D019803:Glutathione Disulfide / Q000378:metabolism; D022781:Hepatocytes / Q000201:enzymology / Q000378:metabolism / Q000502:physiology*; D006579:Heterozygote; D006657:Histones / Q000378:metabolism; D007527:Isoenzymes / Q000235:genetics / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D008969:Molecular Sequence Data; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D054481:Thioredoxin Reductase 1 / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism; D054482:Thioredoxin Reductase 2 / Q000235:genetics / Q000378:metabolism; D013879:Thioredoxins / Q000378:metabolism; D014158:Transcription, Genetic
chemical_list: D006657:Histones; D007527:Isoenzymes; D018809:Proliferating Cell Nuclear Antigen; D013879:Thioredoxins; D054481:Thioredoxin Reductase 1; D054482:Thioredoxin Reductase 2; C516125:Txnrd1 protein, mouse; D005978:Glutathione; D019803:Glutathione Disulfide