Analysis of membrane proteins localizing to the inner nuclear envelope in living cells.

Autor:	Smoyer CJ; Stowers Institute for Medical Research, Kansas City, MO 64110., Katta SS; Stowers Institute for Medical Research, Kansas City, MO 64110., Gardner JM; Stowers Institute for Medical Research, Kansas City, MO 64110., Stoltz L; Stowers Institute for Medical Research, Kansas City, MO 64110., McCroskey S; Stowers Institute for Medical Research, Kansas City, MO 64110., Bradford WD; Stowers Institute for Medical Research, Kansas City, MO 64110., McClain M; Stowers Institute for Medical Research, Kansas City, MO 64110., Smith SE; Stowers Institute for Medical Research, Kansas City, MO 64110., Slaughter BD; Stowers Institute for Medical Research, Kansas City, MO 64110., Unruh JR; Stowers Institute for Medical Research, Kansas City, MO 64110., Jaspersen SL; Stowers Institute for Medical Research, Kansas City, MO 64110 slj@stowers.org.; Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS 66160.
Jazyk:	angličtina
Zdroj:	The Journal of cell biology [J Cell Biol] 2016 Nov 21; Vol. 215 (4), pp. 575-590. Date of Electronic Publication: 2016 Nov 09.
DOI:	10.1083/jcb.201607043
Abstrakt:	Understanding the protein composition of the inner nuclear membrane (INM) is fundamental to elucidating its role in normal nuclear function and in disease; however, few tools exist to examine the INM in living cells, and the INM-specific proteome remains poorly characterized. Here, we adapted split green fluorescent protein (split-GFP) to systematically localize known and predicted integral membrane proteins in Saccharomyces cerevisiae to the INM as opposed to the outer nuclear membrane. Our data suggest that components of the endoplasmic reticulum (ER) as well as other organelles are able to access the INM, particularly if they contain a small extraluminal domain. By pairing split-GFP with fluorescence correlation spectroscopy, we compared the composition of complexes at the INM and ER, finding that at least one is unique: Sbh2, but not Sbh1, has access to the INM. Collectively, our work provides a comprehensive analysis of transmembrane protein localization to the INM and paves the way for further research into INM composition and function. (© 2016 Smoyer et al.)
Databáze:	MEDLINE
Externí odkaz:	Zobrazit plný text záznamu