Multiplexed Barcode Sequencing

Web supplement to
"Highly-multiplexed barcode sequencing: an efficient method for parallel analysis of pooled samples"

Andrew M Smith, Lawrence E Heisler, Robert P St.Onge, Eveline Farias-Hesson, Iain M Wallace, John Bodeau, Adam N. Harris, Kathleen M. Perry, Guri Giaever, Nader Pourmand, Corey Nislow

Nucleic Acids Research, doi:10.1093/nar/gkq368

Supplemental Table 1: List of cisplatin sensitive strains determined by 20-plex barseq analysis

Strains sensitive to 100uM cisplatin were determined by calculation of the fitness defect ratio (control-cisplatin) from barcode sequence counts. Strains with a ratio>1.5 were considered to be sensitive to cisplatin. Strains with a Bar-seq count less than 40 in the DMSO control were excluded. The genes annotation is based on the Saccharomyces Genome Database (SGD, www.yeastgenome.org)

Gene

SGD description

ARC18

Subunit of the ARP2/3 complex, which is required for the motility and integrity of cortical actin patches

ARG1

Arginosuccinate synthetase, catalyzes the formation of L-argininosuccinate from citrulline and L-aspartate in the arginine biosynthesis pathway; potential Cdc28p substrate

ARP4

Nuclear actin-related protein involved in chromatin remodeling, component of chromatin-remodeling enzyme complexes

ATG26

UDP-glucose:sterol glucosyltransferase, conserved enzyme involved in synthesis of sterol glucoside membrane lipids; in contrast to ATG26 from P. pastoris, S. cerevisiae ATG26 is not involved in autophagy

CDC31

Calcium-binding component of the spindle pole body (SPB) half-bridge, required for SPB duplication in mitosis and meiosis II; homolog of mammalian centrin; binds multiubiquitinated proteins and is involved in proteasomal protein degradation

CTS2

Protein similar to Ashbya gossypii sporulation-specific chitinase

CWH41

Processing alpha glucosidase I, ER type II integral membrane N-glycoprotein involved in assembly of cell wall beta 1,6 glucan and asparagine-linked protein glycosylation; also involved in ER protein quality control and sensing of ER stress

HPC2

Subunit of the HIR complex, a nucleosome assembly complex involved in regulation of histone gene transcription; mutants display synthetic defects with subunits of FACT, a complex that allows passage of RNA Pol II through nucleosomes

IOC4

Member of a complex (Isw1b) with Isw1p and Ioc2p that exhibits nucleosome-stimulated ATPase activity and acts within coding regions to coordinate transcription elongation with termination and processing, contains a PWWP motif

LAS21

Integral plasma membrane protein involved in the synthesis of the glycosylphosphatidylinositol (GPI) core structure; mutations affect cell wall integrity

MMS2

Ubiquitin-conjugating enzyme variant involved in error-free postreplication repair; forms a heteromeric complex with Ubc13p, an active ubiquitin-conjugating enzyme; cooperates with chromatin-associated RING finger proteins, Rad18p and Rad5p

OSM1

Fumarate reductase, catalyzes the reduction of fumarate to succinate, required for the reoxidation of intracellular NADH under anaerobic conditions; mutations cause osmotic sensitivity

PCK1

Phosphoenolpyruvate carboxykinase, key enzyme in gluconeogenesis, catalyzes early reaction in carbohydrate biosynthesis, glucose represses transcription and accelerates mRNA degradation, regulated by Mcm1p and Cat8p, located in the cytosol

PSF3

Subunit of the GINS complex (Sld5p, Psf1p, Psf2p, Psf3p), which is localized to DNA replication origins and implicated in assembly of the DNA replication machinery

PSY2

Putative subunit of an evolutionarily conserved protein phosphatase complex containing the catalytic subunit Pph3p and the regulatory subunit Psy4p; required for cisplatin and oxaliplatin resistance; putative homolog of mammalian R3

RAD5

DNA helicase proposed to promote replication fork regression during postreplication repair by template switching; RING finger containing ubiquitin ligase; stimulates the synthesis of free and PCNA-bound polyubiquitin chains by Ubc13p-Mms2p

RAD51

Strand exchange protein, forms a helical filament with DNA that searches for homology; involved in the recombinational repair of double-strand breaks in DNA during vegetative growth and meiosis; homolog of Dmc1p and bacterial RecA protein

RAD10

Single-stranded DNA endonuclease (with Rad1p), cleaves single-stranded DNA during nucleotide excision repair and double-strand break repair; subunit of Nucleotide Excision Repair Factor 1 (NEF1); homolog of human ERCC1 protein

RSC58

Component of the RSC chromatin remodeling complex; RSC functions in transcriptional regulation and elongation, chromosome stability, and establishing sister chromatid cohesion; involved in telomere maintenance

SNF2

Catalytic subunit of the SWI/SNF chromatin remodeling complex involved in transcriptional regulation; contains DNA-stimulated ATPase activity; functions interdependently in transcriptional activation with Snf5p and Snf6p

VID28

Protein involved in proteasome-dependent catabolite degradation of fructose-1,6-bisphosphatase (FBPase); localized to the nucleus and the cytoplasm

YBL081W

Non-essential protein of unknown function; null mutation results in a decrease in plasma membrane electron transport

YEL008W

Hypothetical protein predicted to be involved in metabolism

YER188W

Hypothetical protein

YGR079W

Putative protein of unknown function; YGR079W is not an essential gene

YKU70

Subunit of the telomeric Ku complex (Yku70p-Yku80p), involved in telomere length maintenance, structure and telomere position effect; relocates to sites of double-strand cleavage to promote nonhomologous end joining during DSB repair

YNL187W

Protein involved in mRNA splicing; contains a consensus nuclear export signal (NES) sequence similar to the consensus sequence recognized by Crm1p; interacts genetically with Prp40p and Tgs1p; contains WD40 repeats

YOR199W

Dubious open reading frame unlikely to encode a protein, based on available experimental and comparative sequence data

Multiplexed Barcode SequencingInquiries can be addressed to guri.giaever@utoronto.ca OR corey.nislow@utoronto.ca