title: The core proteins A2 and B1 exist as (A2)3B1 tetramers in 40S nuclear ribonucleoprotein particles.
abstract: The six "core" proteins of HeLa cell 40S nuclear ribonucleoprotein particles (hnRNP particles) package 700-nucleotide lengths of pre-mRNA into a repeating array of regular particles. We have previously shown that the C proteins exist as anisotropic tetramers of (C1)3C2 in 40S hnRNP particles and that each particle probably contains three such tetramers. We report here that proteins A2 and B1 also exist in monoparticles as (A2)3B1 tetramers and that each monoparticle contains at least three such tetramers. Proteins A2 and B1 dissociate from isolated monoparticles as a stable tetramer upon nuclease digestion. In low-salt gradients, the tetramers sediment at 6.8S, which is consistent with a mass of 145 kDa. In 200 mM salt, the concentration which dissociates these proteins from RNA, only 4.2S dimers exist in solution. Tetramers of (A2)3B1 possess the ability to package multiples of 700 nucleotides of RNA in vitro into an array of regular, 22.5-nm 43S particles. Unlike the in vitro assembly of intact 40S hnRNP, the (A2)3B1 tetramers assemble by means of a highly cooperative process. These findings indicate that the (A2)3B1 tetramers play a major role in hnRNP assembly and they further support the contention that 40S monoparticles are regular structures composed of three copies of three different tetramers, i.e., 3[(A1)3B2, (A2)3B1, (C1)3C2].
authors.affiliation: Department of Molecular Biology, Vanderbilt University, Nashville, Tennessee 37235.
authors.forename: S F
authors.initials: SF
authors.lastname: Barnett
authors.forename: T A
authors.initials: TA
authors.lastname: Theiry
authors.forename: W M
authors.initials: WM
authors.lastname: LeStourgeon
mesh_terms: D002467:Cell Nucleus / Q000378:metabolism; D002499:Centrifugation, Density Gradient; D004591:Electrophoresis, Polyacrylamide Gel; D006367:HeLa Cells / Q000378:metabolism; D006801:Humans; D046911:Macromolecular Substances; D008854:Microscopy, Electron; D008970:Molecular Weight; D012261:Ribonucleoproteins / Q000302:isolation & purification* / Q000648:ultrastructure
chemical_list: D046911:Macromolecular Substances; D012261:Ribonucleoproteins