Establishment of an "in saccharo" experimental system.

Genes & Genetic Systems
Tetsushi Iida, Takehiko Kobayashi

Abstract

Many proteins form complexes that function in reaction pathways. Therefore, to understand protein function, it is necessary to reconstitute complexes and pathways in vitro. However, it is not straightforward to achieve full activity in reconstituted systems. To address this problem, we present a yeast system named "in saccharo" analysis, which uses budding yeast for simultaneous expression and analysis of many kinds of non-host proteins, such as human proteins. For this purpose, vectors that can accommodate many genes are required. Here, we describe the construction of a chromosome vector by insertion of unique barcode sequences (BCs) into the ribosomal RNA gene repeat (rDNA). Each unit of the rDNA has a BC that is used as the target for integration of an external gene. Because rDNA is naturally capable of maintaining many repetitive copies, the vector is expected to retain the numerous external genes that may be required for reconstitution of functional protein complexes and reaction pathways. Consistent with this prediction, we were able to clone three human genes that form the RNA silencing pathway, which has no functional equivalent in budding yeast, and to demonstrate functionality in this in saccharo analysis system.

References

Mar 20, 1985·Journal of Molecular Biology·H ArakiY Oshima
Sep 15, 1983·Nature·A W Murray, J W Szostak
May 1, 1984·Molecular and Cellular Biology·J H ProffittS Hattman
Jan 1, 1980·Annual Review of Biochemistry·E O Long, I B Dawid
May 1, 1994·The Journal of Cell Biology·V GuacciD Koshland
May 2, 1997·Science·M K RaghuramanW L Fangman
Jul 12, 2002·Nature·Gregory J Hannon
Nov 8, 2005·Cell·Richard I GregoryRamin Shiekhattar
Oct 24, 2007·Genome Biology·Jason E StajichScott W Roy
Jan 8, 2008·Proceedings of the National Academy of Sciences of the United States of America·Ian J MacRaeJennifer A Doudna
Sep 12, 2009·Science·Ines A DrinnenbergDavid P Bartel
Nov 10, 2009·Biochemistry and Cell Biology = Biochimie Et Biologie Cellulaire·Benjamin R HarrisonJocelyn E Krebs
Jan 22, 2011·Nucleic Acids Research·Kyoungho SukFrederick P Roth
Apr 27, 2011·Genes to Cells : Devoted to Molecular & Cellular Mechanisms·Takaaki Miyazaki, Takehiko Kobayashi
Nov 19, 2013·Proteomics·Trevor Clancy, Eivind Hovig
Apr 15, 2014·Proceedings of the Japan Academy. Series B, Physical and Biological Sciences·Takehiko Kobayashi
Jul 23, 2014·Philosophical Transactions of the Royal Society of London. Series B, Biological Sciences·John V Kilmartin
Mar 11, 2017·Science·Sarah M RichardsonJoel S Bader
Dec 5, 2017·Cell·David M Truong, Jef D Boeke
Jan 8, 2019·Molecular Cell·Tetsushi Iida, Takehiko Kobayashi

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