Degradation signals within both terminal domains of the cauliflower mosaic virus capsid protein precursor.
Targeted protein degradation plays an important regulatory role in the cell, but only a few protein degradation signals have been characterized in plants. Here we describe three instability determinants in the termini of the cauliflower mosaic virus (CaMV) capsid protein precursor, of which one is still present in the mature capsid protein p44. A modified ubiquitin protein reference technique was used to show that these motifs are still active when fused to a heterologous reporter gene. The N-terminus of p44 contains a degradation motif characterized by proline, glutamate, aspartate, serine and threonine residues (PEST), which can be inactivated by mutation of three glutamic acid residues to alanines. The signals from the precursor do not correspond to known degradation motifs, although they confer high instability on proteins expressed in plant protoplasts. All three instability determinants were also active in mammalian cells. The PEST signal had a significantly higher degradation activity in HeLa cells, whereas the precursor signals were less active. Inhibition studies suggest that only the signal within the N-terminus of the precursor is targeting the proteasome in plants. This implies that the other two signals may target a novel degradation pathway.
Karsies
A
A
Friedrich-Miescher Institute, CH-4002 Basel, Switzerland.
Hohn
T
T
Leclerc
D
D
D000595:Amino Acid Sequence; D002213:Capsid / Q000737:chemistry / Q000378:metabolism*; D017796:Caulimovirus / Q000378:metabolism*; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics; D003546:Cysteine Endopeptidases / Q000378:metabolism; D005966:Glucuronidase / Q000235:genetics; D006868:Hydrolysis; D008969:Molecular Sequence Data; D009097:Multienzyme Complexes / Q000378:metabolism; D046988:Proteasome Endopeptidase Complex; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D015398:Signal Transduction
D009097:Multienzyme Complexes; D011498:Protein Precursors; D011993:Recombinant Fusion Proteins; D015500:Chloramphenicol O-Acetyltransferase; D005966:Glucuronidase; D003546:Cysteine Endopeptidases; D046988:Proteasome Endopeptidase Complex