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JCS ePress
online publication date 13 May 2008
doi: 10.1242/jcs.020321
Research Article
Distinct targeting pathways for the membrane insertion of tail-anchored (TA) proteins
Vincenzo Favaloro,
Milan Spasic,
Blanche Schwappach,
and
Bernhard Dobberstein*
* Author for correspondence (e-mail: b.dobberstein{at}zmbh.uni-heidelberg.de)
Tail-anchored (TA) proteins are characterised by a C-terminal transmembrane region that mediates post-translational insertion into the membrane of the endoplasmic reticulum (ER). We have investigated the requirements for membrane insertion of three TA proteins, RAMP4, Sec61
and cytochrome b5. We show here that newly synthesised RAMP4 and Sec61
can accumulate in a cytosolic, soluble complex with the ATPase Asna1 before insertion into ER-derived membranes. Membrane insertion of these TA proteins is stimulated by ATP, sensitive to redox conditions and blocked by alkylation of SH groups by N-ethylmaleimide (NEM). By contrast, membrane insertion of cytochrome b5 is not found to be mediated by Asna1, not stimulated by ATP and not affected by NEM or an oxidative environment. The Asna1-mediated pathway of membrane insertion of RAMP4 and Sec61
may relate to functions of these proteins in the ER stress response.

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