ATP induces large quaternary rearrangements in a cage-like chaperonin structure.

BACKGROUND: The chaperonins, a family of molecular chaperones, are large oligomeric proteins that bind nonnative intermediates of protein folding. They couple the release and correct folding of their ligands to the binding and hydrolysis of ATP. Chaperonin 60 (cpn60) is a decatetramer (14-mer) of 60 kD subunits. Folding of some ligands also requires the cooperation of cpn10, a heptamer of 10 kD subunits. RESULTS: We have determined the three-dimensional arrangements of subunits in Rhodobacter sphaeroides cpn60 in the nucleotide-free and ATP-bound forms. Negative stain electron microscopy and tilt reconstruction show the cylindrical structure of the decatetramer comprising two rings of seven subunits. The decatetramer consists of two cages joined base-to-base without a continuous central channel. These cages appear to contain bound polypeptide with an asymmetric distribution between the two rings. The two major domains of each subunit are connected on the exterior of the cylinder by a narrower bridge of density that could be a hinge region. Binding of ATP to cpn60 causes a major rearrangement of the protein density, which is reversed upon the hydrolysis of the ATP. Cpn10 binds to only one end of the cpn60 structure and is visible as an additional layer of density forming a cap on one end of the cpn60 cylinder. CONCLUSIONS: The observed rearrangement is consistent with an inward 5-10 degrees rotation of subunits, pivoting about the subunit contacts between the two heptamers, and thus bringing cpn60 domains towards the position occupied by the bound polypeptide. This change could explain the stimulation of ATPase activity by ligands, and the effects of ATP on lowering the affinity of cpn60 for ligands and on triggering the release of folding polypeptides.

Photoreceptors controlling transcription of rbcS genes in green leaf tissue of Pisum sativum.

We have investigated the photoreceptors controlling transcription of genes encoding the small subunit (rbcS) of ribulose 1,5-bisphosphate carboxylase-oxygenase in green leaf tissue of pea (Pisum sativum). RbcS transcription was measured by hybridising labelled transcripts of isolated nuclei to rbcS cDNA clones. Transfer of green Pisum leaf tissue to darkness for 5 h causes a substantial decrease in the rate of rbcS transcription and the rate is restored rapidly when the plants are returned to white light. Low fluence rates of red light are ineffective in restoring the rate of rbcS transcription, suggesting that phytochrome alone does not fully mediate the response. Blue light is similarly effective to white light of an equal fluence rate (120 mumol m-2 s-1) in restoring the rate of rbcS transcription in the dark-treated plants, indicating that a blue light photoreceptor is involved. However, red light at the same fluence rate produces about 65% of the effect of blue or white light, showing that the blue light photoreceptor is not the only photoreceptor controlling rbcS transcription in the green leaf tissue. The identity of the photoreceptor responsible for the red light effect is discussed. Similar effects of blue and red light are observed at the level of transcript abundance in dark-grown pea leaf tissue given a brief illumination with red light, which potentiates the tissue for rapid transcript accumulation in white light.

Genetic differences between inbred strains of mice; a new source of variation in high-sulphur keratins.

Proteins were extracted by improved techniques from the hair of inbred strains of Peru, C57BL/6 and CBA/Ca mice. The extracts were characterized by amino acid analysis and high resolution polyacrylamide gel electrophoresis, and previously unreported strain differences were observed. Genetic analysis confirmed them.