Illuminating the human genome

The identification and analysis of novel genes and their encoded protein products remains a vigorous area of research in biology today. Worldwide genomic and cDNA sequencing projects are now identifying new molecules every day and the need for methodologies to functionally characterise these proteins has never been greater. The distinct compartmental arrangement of eukaryotic cells helps define the processes which occur within or in proximity to these membranes, and as such provides one means of inferring protein function. We describe here some of the methods recently reported in the literature, which use the subcellular localisation of proteins as a first step towards their further characterisation.

[1]  P. Bastiaens,et al.  Fluorescence lifetime imaging microscopy: spatial resolution of biochemical processes in the cell. , 1999, Trends in cell biology.

[2]  A. Poustka,et al.  Systematic subcellular localization of novel proteins identified by large‐scale cDNA sequencing , 2000, EMBO reports.

[3]  C. González,et al.  Protein traps: using intracellular localization for cloning. , 2000, Trends in cell biology.

[4]  P. Nurse,et al.  Identification of fission yeast nuclear markers using random polypeptide fusions with green fluorescent protein. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[5]  D. Eisenberg,et al.  Localizing proteins in the cell from their phylogenetic profiles. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[6]  R Pepperkok,et al.  COPI-coated ER-to-Golgi transport complexes segregate from COPII in close proximity to ER exit sites. , 2000, Journal of cell science.

[7]  R. Pepperkok,et al.  Observing proteins in their natural habitat: the living cell. , 2000, Trends in biochemical sciences.

[8]  E. Stelzer,et al.  Photobleaching GFP reveals protein dynamics inside live cells. , 1999, Trends in cell biology.

[9]  Jyoti S. Choudhary,et al.  Proteomics Characterization of Abundant Golgi Membrane Proteins* , 2001, The Journal of Biological Chemistry.

[10]  André Goffeau,et al.  The yeast genome directory. , 1997, Nature.

[11]  S. Cutler,et al.  Random GFP::cDNA fusions enable visualization of subcellular structures in cells of Arabidopsis at a high frequency. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[12]  J. Yates,et al.  Proteomic Analysis of Two Functional States of the Golgi Complex in Mammary Epithelial Cells , 2000, Traffic.

[13]  A. Matus,et al.  GFP illuminates the cytoskeleton. , 1998, Trends in cell biology.

[14]  S Tweedie,et al.  Capturing novel mouse genes encoding chromosomal and other nuclear proteins. , 1998, Journal of cell science.

[15]  R. Singer,et al.  Determinants of mRNA localization. , 1992, Current opinion in cell biology.

[16]  Stephen M. Mount,et al.  The genome sequence of Drosophila melanogaster. , 2000, Science.

[17]  L. Huber,et al.  Subcellular fractionation, electromigration analysis and mapping of organelles. , 1999, Journal of chromatography. B, Biomedical sciences and applications.

[18]  C. González,et al.  Motif trap: a rapid method to clone motifs that can target proteins to defined subcellular localisations. , 1999, Journal of cell science.

[19]  P Bork,et al.  Wanted: subcellular localization of proteins based on sequence. , 1998, Trends in cell biology.

[20]  Stephen S. Taylor,et al.  A Visual Screen of a Gfp-Fusion Library Identifies a New Type of Nuclear Envelope Membrane Protein , 1999, The Journal of cell biology.

[21]  Rainer Pepperkok,et al.  Visualization of ER-to-Golgi Transport in Living Cells Reveals a Sequential Mode of Action for COPII and COPI , 1997, Cell.

[22]  R Y Tsien,et al.  Understanding, improving and using green fluorescent proteins. , 1995, Trends in biochemical sciences.

[23]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[24]  T Nakahata,et al.  A method to identify cDNAs based on localization of green fluorescent protein fusion products. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[25]  Roger Y. Tsien,et al.  Crystal Structure of the Aequorea victoria Green Fluorescent Protein , 1996, Science.

[26]  J. Lippincott-Schwartz,et al.  Dual-colour imaging with GFP variants. , 1999, Trends in cell biology.

[27]  S. Lukyanov,et al.  Fluorescent proteins from nonbioluminescent Anthozoa species , 1999, Nature Biotechnology.

[28]  J. Berg Genome sequence of the nematode C. elegans: a platform for investigating biology. , 1998, Science.