真菌子实体的形成需要.ppt

Absence of CsnA or CsnD Prevents the Assembly of CsnE with Other CSN Subunits. The JAMM Motif of CsnE Is Essential for Fungal Fruit Body Formation. JAMM domain proteins CSN subunit 5, 26S proteasome LID subunit RPN11/PSMD14, and AMSH of A. nidulans (an), H. sapiens (hs), A. thaliana (at), S. pombe (sp), or S. cerevisiae (sc) were aligned by ClustalW with the archaea JAMM protein AF2198. JAMM is highlighted in black, and additional conserved residues are red (100%) or green (40%). Residues mutated in JAMM (gray bar) are marked by asterisks csnE1 (D147N) and csnE2 (H134A, H136A, and D147N) were transcribed as shown by Northern hybridization csnE1 and csnE2 did not rescue the csnE mutant phenotypes…… 串联亲和纯化------TAP DISSCUSSION The complex is essential for development,and defects in CSN result in embryonic death of these multicellular organisms The fifth subunit (CsnE/Jab1) of CSN includes the only known enzyme activity of the complex, a metalloprotease that binds zinc ions by its conserved JAMM motif. In the fungal system, CsnE/Jab1 is unable to recruit any other CSN subunit and is less stable when CSN subunits 1 or 4 are absent. This argues against stable CsnE subcomplexes in fungi, although it does not exclude the formation of CsnE oligomers or unstable CsnE-containing subcomplexes. The A. nidulans genome reveals that all major genes for putative substrates of the CSN deneddylase activity are present. These include a proposed Nedd8 encoding sequence and the three putative cullins Cul1 , Cul3 , und Cul4 . Cullins are part of cullin-RING E3 ligase cores and are known to assemble with various specific substrate-adaptors like F-box proteins the control of specific E3 ubiquitin ligases by CSN seems to be less critical for hyphal growth itself than for fungal developmental processes where hyphae have to be reconstructed to allow the maturation of fruit bodies. This suggests that the importance of a stringent control of ubiquitin-dependent protein degradation is less