Jeffrey A. Hussmann

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Ribosome-footprint profiling provides genome-wide snapshots of translation, but technical challenges can confound its analysis. Here, we use improved methods to obtain ribosome-footprint profiles and mRNA abundances that more faithfully reflect gene expression in Saccharomyces cerevisiae. Our results support proposals that both the beginning of coding(More)
Ribosome profiling produces snapshots of the locations of actively translating ribosomes on messenger RNAs. These snapshots can be used to make inferences about translation dynamics. Recent ribosome profiling studies in yeast, however, have reached contradictory conclusions regarding the average translation rate of each codon. Some experiments have used(More)
A major limitation of high-throughput DNA sequencing is the high rate of erroneous base calls produced. For instance, Illumina sequencing machines produce errors at a rate of ~0.1-1 × 10(-2) per base sequenced. These technologies typically produce billions of base calls per experiment, translating to millions of errors. We have developed a unique library(More)
CRISPR-Cas nucleoproteins target foreign DNA via base pairing with a crRNA. However, a quantitative description of protein binding and nuclease activation at off-target DNA sequences remains elusive. Here, we describe a chip-hybridized association-mapping platform (CHAMP) that repurposes next-generation sequencing chips to simultaneously measure the(More)
Ribosome stalling leads to recruitment of the ribosome quality control complex (RQC), which targets the partially synthesized polypeptide for proteasomal degradation through the action of the ubiquitin ligase Ltn1p. A second core RQC component, Rqc2p, modifies the nascent polypeptide by adding a carboxyl-terminal alanine and threonine (CAT) tail through a(More)
It has been proposed that patterns in the usage of synonymous codons provide evidence that individual tRNA molecules are recycled through the ribosome, translating several occurrences of the same amino acid before diffusing away. The claimed evidence is based on counting the frequency with which pairs of synonymous codons are used at nearby occurrences of(More)
Schmitt et al. (1) raise the concern that circle sequencing (2) may spuriously count information from the same starting molecule multiple times. There are indeed several mechanisms by which multiple final reads produced by the circle-sequencing process could be derived from the same starting molecule, as discussed briefly in the last paragraph of our(More)
Ribosomes contain proteins that must themselves be made by ribosomes. A new study shows that splitting ribosomal protein content into many small, similarly sized units maximizes the efficiency of this synthesis, suggesting that ribosomal architecture has been shaped by evolutionary pressure to efficiently self-synthesize.
All ribosome profiling experiments analyzed involve attaching a known sequence to the 3′ end of RNA footprints to which a reverse transcription primer can be annealed. Some experiments use polyA tailing for this purpose, while others attach an oligonucleotide linker sequence. For experiments using polyA tailing, reads were trimmed from the end back to the(More)
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