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S M Gasser

Publications and source records attributed to S M Gasser.

96 records · Page 6Linked to original sources
PubMed1982

Import of proteins into mitochondria. Energy-dependent, two-step processing of the intermembrane space enzyme cytochrome b2 by isolated yeast mitochondria.

Import of in vitro-synthesized cytochrome b2 (a soluble intermembrane space enzyme) was studied wih isolated yeast mitochondria. Import requires an electrochemical gradient across the inner membrane and is accompanied by cleavage of the precursor to the corresponding mature form. This conversion proceeds via an intermediate form of cytochrome b2, which can be detected as a transient species when mitochondria are incubated with the cytochrome b2 precursor for short times or at low temperatures. Conversion of the precursor to the intermediate form is energy-dependent and catalyzed by an o-phenanthroline-sensitive protease located in the soluble matrix. The intermediate is subsequently converted to mature cytochrome b2 in a reaction which is o-phenanthroline-insensitive and requires neither an energized inner membrane nor a soluble component of the intermembrane space. Whereas mature cytochrome b2 is soluble, the intermediate formed by isolated mitochondria is membrane-bound and exposed to the intermembrane space. The same intermediate is detected as a transient species during cytochrome b2 maturation in intact yeast cells (Reid, G. A., Yonetani, T., and Schatz, G (1982) J. Biol. Chem. 257, 13068-13074). The in vitro studies reported here suggest that a part of the cytochrome b2 precursor polypeptide chain is transported to the matrix where it is cleaved to a membrane-bound intermediate form by the same protease that processes polypeptides destined for the matrix space or for the inner membrane. In a second reaction, the cytochrome b2 intermediate is converted to mature cytochrome b2 which is released into the intermembrane space. The binding of heme is not necessary for converting the intermediate to the mature polypeptide.

Biological Transport↗
PubMed1982

Imported mitochondrial proteins cytochrome b2 and cytochrome c1 are processed in two steps.

Cytochrome b2 of yeast is located in the space between the inner and outer mitochondrial membranes whereas cytochrome c1 is bound to the outer face of the mitochondrial inner membrane. Both proteins are made outside the mitochondria as larger precursors that are processed to their mature forms in two steps. In the first step, at least a segment of the precursor polypeptide chain penetrates into the mitochondrial matrix and is cleaved to an intermediate form by a matrix-localized soluble protease. This step requires an electrochemical gradient across the mitochondrial inner membrane. In the second step, the intermediate form is converted to the mature form. For cytochrome c1, this second step requires heme. Import of these two cytochromes thus involves a "detour" across the inner membrane.

Biological Transport↗
PubMedDate not supplied

Imaging the asymmetrical DNA bend induced by repressor activator protein 1 with scanning tunneling microscopy.

The yeast Repressor Activator Protein 1 (RAP1) binds a 13-bp consensus found in many transcriptional regulatory regions, in silencer elements, and in telomeric repeat DNA of Saccharomyces cerevisiae. Gel retardation assays suggest that RAP1 bends DNA as it binds, with the vertex of the angle located 5' of the consensus. We show that removal of 230 aa in the N-terminus of RAP1 reduces the aberrant electrophoretic mobility of the protein-DNA complex, while removal of a C-terminal domain of RAP1 causes even greater distortion. To demonstrate that the aberrant electrophoretic mobility is really due to a bend in the double helix, the RAP1-DNA complex was analyzed by Scanning Tunnelling Microscopy (STM). The efficiency and accuracy of binding is checked in parallel by standard Transmission Electron Microscopy (TEM). Due to the use of high-angle shadowing of freeze-dried samples at low temperatures, the STM images allow us to confirm that RAP1 binding induces a DNA bend > 50 degrees, while the binding of the minimal DNA-binding domain shows significantly less distortion of the DNA helix.

DNA↗
PubMedDate not supplied

The nucleolus: nucleolar space for RENT.

Recent studies indicate that the nucleolus is not just a site of ribosome biogenesis. Intriguing links have been found between nucleolar components and the machinery that regulates the cell cycle.

Cell Cycle↗