October 18, 2012

Reverse translocation sends proteins to the cytosol for degradation

  • Retrograde translocation (Reverse translocation) is the translocation of a protein from the lumen of the ER to the cytoplasm. It usually occurs to allow misfolded or damaged proteins to be degraded by the proteasome.
  • Sec61 translocons can be used for reverse translocation of proteins from the ER into the cytosol. 

Several important activities occur within the endoplasmic reticulum. Proteins move through the ER en route to a variety of destinations (see 27 Protein trafficking). They are glycosylated and folded into their final conformations. The ER provides a "quality control" system in which misfolded proteins are identified and degraded. However, the degradation itself does not occur in the ER, but may require the protein to be exported back to the cytosol.
The first indication that ER proteins are degraded in the cytosol and not in the ER itself was provided by evidence for the involvement of the proteasome, a large protein aggregate with several proteolytic activities (see 8.32 The proteasome is a large machine that degrades ubiquitinated proteins). Inhibitors of the proteasome prevent the degradation of aberrant ER proteins. Proteins are marked for cleavage by the proteasome when they are modified by the addition of ubiquitin, a small polypeptide chain (see 8.31 Ubiquitination targets proteins for degradation). The important point to note now is that ubiquitination and proteasomal degradation both occur in the cytosol (with a minor proportion in the nucleus).
Transport from the ER back into the cytosol occurs by a reversal of the usual process of import (for review see Tsai, Ye, and Rapoport, 2002). This is called reverse translocation. The Sec61 translocon is used. The conditions are different; for example, the translocon is not associated with a ribosome. Some mutations in Sec61 prevent reverse translocation, but do not prevent forward translocation (Zhou and Schekman, 1999; Wilkinson et al., 2000). This could be either because there is some difference in the process or (more likely) because these regions interact with other components that are necessary for reverse translocation.

Figure 8.30 points out that we do not know how the channel is opened to allow insertion of the protein on the ER side. Special components are presumably involved. One model is that misfolded or misassembled proteins are recognized by chaperones, which transfer them to the translocon (for review see Johnson and Haigh, 2000). In one particular case, human cytomegalovirus (CMV) codes for cytosolic proteins that destroy newly synthesized MHC class I (cellular major histocompatibility complex) proteins. This requires a viral protein product (US2), which is a membrane protein that functions in the ER. It interacts with the MHC proteins and probably conveys them into the translocon for reverse translocation.
The system involved in the degradation of aberrant ER proteins can be identified by mutations (in yeast) that lead to accumulation of aberrant proteins. Usually a protein that misfolds (produced by a mutated gene) is degraded instead of being transported through the ER. Yeast mutants that cannot degrade the substrate fall into two classes: some identify components of the proteolytic apparatus, such as the enzymes involved in ubiquitination; other identify components of the transport apparatus, including Sec61, BiP, and Sec63. There is also a protein in the ER membrane that functions on the cytosolic side to localize the ubiquitination enzymes at the translocon. In fact, retrograde transport into the cytosol cannot occur in the absence of this protein, which suggests that there is a mechanical link between retrograde transport and degradation (Wiertz et al., 1996).