Biomedical subjects

Jeremy D Brown

Publications and source records attributed to Jeremy D Brown.

5 recordsLinked to original sources

PubMed2005

A nomenclature for all signal recognition particle RNAs.

The signal recognition particle (SRP) is a cytosolic ribonucleoprotein complex that guides secretory proteins to biological membranes in all organisms. The SRP RNA is at the center of the structure and function of the SRP. The comparison of the growing number of SRP RNA sequences provides a rich source for gaining valuable insight into the composition, assembly, and phylogeny of the SRP. In order to assist in the continuation of these studies, we propose an SRP RNA nomenclature applicable to the three divisions of life.

Animals↗

PubMed2004

Saccharomyces SRP RNA secondary structures: a conserved S-domain and extended Alu-domain.

The contribution made by the RNA component of signal recognition particle (SRP) to its function in protein targeting is poorly understood. We have generated a complete secondary structure for Saccharomyces cerevisiae SRP RNA, scR1. The structure conforms to that of other eukaryotic SRP RNAs. It is rod-shaped with, at opposite ends, binding sites for proteins required for the SRP functions of signal sequence recognition (S-domain) and translational elongation arrest (Alu-domain). Micrococcal nuclease digestion of purified S. cerevisiae SRP separated the S-domain of the RNA from the Alu-domain as a discrete fragment. The Alu-domain resolved into several stable fragments indicating a compact structure. Comparison of scR1 with SRP RNAs of five yeast species related to S. cerevisiae revealed the S-domain to be the most conserved region of the RNA. Extending data from nuclease digestion with phylogenetic comparison, we built the secondary structure model for scR1. The Alu-domain contains large extensions, including a sequence with hallmarks of an expansion segment. Evolutionarily conserved bases are placed in the Alu- and S-domains as in other SRP RNAs, the exception being an unusual GU(4)A loop closing the helix onto which the signal sequence binding Srp54p assembles (domain IV). Surprisingly, several mutations within the predicted Srp54p binding site failed to disrupt SRP function in vivo. However, the strength of the Srp54p-scR1 and, to a lesser extent, Sec65p-scR1 interaction was decreased in these mutant particles. The availability of a secondary structure for scR1 will facilitate interpretation of data from genetic analysis of the RNA.

Alu Elements↗

PubMed2003

Co-translational, intraribosomal cleavage of polypeptides by the foot-and-mouth disease virus 2A peptide.

During co-translational protein import into the endoplasmic reticulum ribosomes are docked onto the translocon. This prevents inappropriate exposure of nascent chains to the cytosol and, conversely, cytosolic factors from gaining access to the nascent chain. We exploited this property of co-translational translocation to examine the mechanism of polypeptide cleavage by the 2A peptide of the foot-and-mouth disease virus. We find that the scission reaction is unaffected by placing 2A into a co-translationally targeted protein. Moreover, the portion of the polypeptide C-terminal to the cleavage site remains in the cytosol unless it contains its own signal sequence. The pattern of cleavage is consistent with the proposal that the 2A-mediated cleavage reaction occurs within the ribosome itself. In addition, our data indicate that the ribosome-translocon complex detects the break in the nascent chain and prevents any downstream protein lacking a signal sequence from gaining access to the endoplasmic reticulum.

Endoplasmic Reticulum↗

PubMed2003

Protein A-tagging for purification of native macromolecular complexes from Candida albicans.

Protein A-tagging has become an important tool in characterization of protein-protein interactions in many systems, allowing purification of multicomponent complexes under native conditions. Here we provide a set of vectors that allow protein A-tagging in Candida albicans, through addition of the tag to open reading frames. These vectors were successfully used to generate stably tagged proteins that were functional, shown to be localized appropriately or assembled into complexes. These new vectors comprise a useful addition to the C. albicans molecular toolbox.

Base Sequence↗

PubMedDate not supplied

[Non-canonical decoding events at stop codons in eukaryotes].

Regulation of protein synthesis at translation termination is a relatively under-explored, but rapidly expanding field. Recent advances in elucidating the mechanism of translation termination are helping to understand non-canonical events associated with translation termination. These "recoding" events include read-through of stop-codons, insertion of unusual amino acids such as selenocysteine and production of several polypeptides from one open reading frame. This review summarises data on termination-dependent recoding events, and proposes that there are two types of stop codon-associated sequences optimized to perform different functions: termination of translation per se or alternative elongation events.

Animals↗