Localization of proteins that are coordinately expressed with Cln2 during the cell cycle

The localization of proteins can give important clues about their function and help sort data from large‐scale proteomic screens. Forty‐five proteins were tagged with the GFP variant YFP. These proteins were chosen because they are encoded by genes that display strong cell cycle‐dependent expression that peaks in G1. Most of these proteins localize to either the nucleus or to sites of cell growth. We are able to assign new cellular component GO terms to ASF2, TOS4, RTT109, YBR070C, YKR090W, YOL007C, YOL019W and YPR174C. We also have localization data for 21 other proteins. Noteworthy localizations were found for Rfa1p, a member of the DNA replication A complex, and Pri2p and Pol12p, subunits of the α‐DNA polymerase : primase complex. In addition to its nuclear localization, Rfa1p assembled into cytoplasmic foci adjacent to the nucleus in cells during the G1–S phase transition of the cell cycle. Pri2 and Pol12 took on a beaded appearance at the G1–S transition and later in the cell cycle were enriched in the nuclear envelope. A new spindle pole body/nuclear envelope component encoded by YPR174 was identified. The cell cycle‐dependent abundance of Tos4p mirrored Yox1p and these two proteins were the only proteins that were found exclusively at the G1–S phase of the cell cycle. A complete list of localizations, along with images, can be found at our website (http://www.yeastrc.org/cln2/). Copyright © 2004 John Wiley & Sons, Ltd.

[1]  G. Fink,et al.  Methods in yeast genetics , 1979 .

[2]  M. Chalfie.  GREEN FLUORESCENT PROTEIN , 1995, Photochemistry and photobiology.

[3]  J. Hegemann,et al.  Green fluorescent protein as a marker for gene expression and subcellular localization in budding yeast , 1996, Yeast.

[4]  R. Sternglanz,et al.  Two New S‐Phase‐Specific Genes from Saccharomyces cerevisiae , 1997, Yeast.

[5]  P. Philippsen,et al.  Heterologous HIS3 Marker and GFP Reporter Modules for PCR‐Targeting in Saccharomyces cerevisiae , 1997, Yeast.

[6]  M. Wold.  Replication protein A: a heterotrimeric, single-stranded DNA-binding protein required for eukaryotic DNA metabolism. , 1997, Annual review of biochemistry.

[7]  P. Burgers,et al.  Eukaryotic DNA polymerases in DNA replication and DNA repair , 1998, Chromosoma.

[8]  Ronald W. Davis,et al.  A genome-wide transcriptional analysis of the mitotic cell cycle. , 1998, Molecular cell.

[9]  R. Tsien,et al.  green fluorescent protein , 2020, Catalysis from A to Z.

[10]  Michael Ruogu Zhang,et al.  Comprehensive identification of cell cycle-regulated genes of the yeast Saccharomyces cerevisiae by microarray hybridization. , 1998, Molecular biology of the cell.

[11]  D. Botstein,et al.  Cluster analysis and display of genome-wide expression patterns. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[12]  A. Kikuchi,et al.  NBP1 (Nap1 binding protein 1), an essential gene for G2/M transition of Saccharomyces cerevisiae, encodes a protein of distinct sub-nuclear localization. , 2000, Gene.

[13]  Monika Tsai-Pflugfelder,et al.  The dynamics of yeast telomeres and silencing proteins through the cell cycle. , 2000, Journal of structural biology.

[14]  B. Kennedy,et al.  Nuclear organization of DNA replication in primary mammalian cells. , 2000, Genes & development.

[15]  A. Shevchenko,et al.  The Bbp1p–Mps2p complex connects the SPB to the nuclear envelope and is essential for SPB duplication , 2000, The EMBO journal.

[16]  M. Johnston,et al.  The yeast genome: on the road to the Golden Age. , 2000, Current opinion in genetics & development.

[17]  L. P. Zhao,et al.  Statistical modeling of large microarray data sets to identify stimulus-response profiles , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[18]  L. Breeden,et al.  Conserved homeodomain proteins interact with MADS box protein Mcm1 to restrict ECB-dependent transcription to the M/G1 phase of the cell cycle. , 2002, Genes & development.

[19]  Lani F. Wu,et al.  Large-scale prediction of Saccharomyces cerevisiae gene function using overlapping transcriptional clusters , 2002, Nature Genetics.

[20]  M. Gerstein,et al.  Complex transcriptional circuitry at the G1/S transition in Saccharomyces cerevisiae. , 2002, Genes & development.

[21]  Yaniv Ziv,et al.  Revealing modular organization in the yeast transcriptional network , 2002, Nature Genetics.

[22]  T. Davis,et al.  Fluorescence resonance energy transfer using color variants of green fluorescent protein. , 2002, Methods in enzymology.

[23]  Ker-Chau Li,et al.  Genome-wide coexpression dynamics: Theory and application , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[24]  C. Ball,et al.  Saccharomyces Genome Database. , 2002, Methods in enzymology.

[25]  Ronald W Davis,et al.  Parallel phenotypic analysis of sporulation and postgermination growth in Saccharomyces cerevisiae , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[26]  Brian E Snydsman,et al.  Assigning function to yeast proteins by integration of technologies. , 2003, Molecular cell.

[27]  E. O’Shea,et al.  Global analysis of protein localization in budding yeast , 2003, Nature.

[28]  Daphne Koller,et al.  Decomposing Gene Expression into Cellular Processes , 2002, Pacific Symposium on Biocomputing.