Different roles of P1 and P2 Saccharomyces cerevisiae ribosomal stalk proteins revealed by cross-linking.
The stalk is an essential domain of the large ribosomal subunit formed by a complex of a set of very acidic proteins bound to a core rRNA binding component. While in prokaryotes there is only one type acidic protein, L7/12, two protein families are found in eukaryotes, phosphoproteins P1 and P2, which presumably have different roles. To search for differences zero-length cross-linking by S-S bridge formation was applied using Saccharomyces cerevisiae mutant P1 and P2 proteins carrying single cysteine residues at various positions. The results show a more exposed location of the N-terminal domain of the P2 proteins, which in contrast to P1, can be found as dimers when the Cys is introduced in this domain. Similarly, the Cys containing C-terminal domain of mutant P2 proteins shows a notable capacity to form cross-links with other proteins, which is considerably lower in the P1 type. On the other hand, mutation at the conserved C-domain of protein P0, the eukaryotic stalk rRNA binding component, results in removal of about 14 terminal amino acids. Protein P2, but not P1, protects mutant P0 from this truncation. These results support a eukaryotic stalk structure in which P1 proteins are internally located with their C-terminals having a restricted reactivity while P2 proteins are more external and accessible to interact with other cellular components.
Qiu
Deyi
D
Centro de Biología Molecular, C.S.I.C. and U.A.M., Canto Blanco, Madrid 28049, Spain.
Parada
Pilar
P
Marcos
Alberto García
AG
Cárdenas
David
D
Remacha
Miguel
M
Ballesta
Juan P G
JP
D003545:Cysteine / Q000737:chemistry; D010750:Phosphoproteins / Q000737:chemistry / Q000502:physiology*; D017434:Protein Structure, Tertiary; D012269:Ribosomal Proteins / Q000737:chemistry / Q000502:physiology*; D012270:Ribosomes / Q000737:chemistry / Q000502:physiology; D012441:Saccharomyces cerevisiae / Q000737:chemistry / Q000502:physiology*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry / Q000502:physiology*
D010750:Phosphoproteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; D003545:Cysteine