The amino terminal end determines the stability and assembling capacity of eukaryotic ribosomal stalk proteins P1 and P2.
The eukaryotic ribosomal proteins P1 and P2 bind to protein P0 through their N-terminal domain to form the essential ribosomal stalk. A mutational analysis points to amino acids at positions 2 and 3 as determinants for the drastic difference of Saccharomyces cerevisiae P1 and P2 half-life, and suggest different degradation mechanisms for each protein type. Moreover, the capacity to form P1/P2 heterodimers is drastically affected by mutations in the P2β four initial amino acids, while these mutations have no effect on P1β. Binding of P2β and, to a lesser extent, P1β to the ribosome is also seriously affected showing the high relevance of the amino acids in the first turn of the NTD α-helix 1 for the stalk assembly. The negative effect of some mutations on ribosome binding can be reversed by the presence of the second P1/P2 couple in the ribosome, indicating a stabilizing structural influence between the two heterodimers. Unexpectedly, some mutations totally abolish heterodimer formation but allow significant ribosome binding and, therefore, a previous P1 and P2 association seems not to be an absolute requirement for stalk assembly. Homology modeling of the protein complexes suggests that the mutated residues can affect the overall protein conformation.
Centro de Biología Molecular Severo Ochoa, Consejo Superior de Investigaciones Científicas and Universidad Autónoma de Madrid, Cantoblanco, 28049 Madrid, Spain.
Hendricka
H
Camargo
Gretel
G
Nusspaumer
David
D
Abia
Verónica
V
Briceño
Miguel
M
Remacha
Juan P G
JP
Ballesta
D019281:Dimerization; D008958:Models, Molecular; D009154:Mutation; D010750:Phosphoproteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D055503:Protein Multimerization; D055550:Protein Stability; D012269:Ribosomal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012270:Ribosomes / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
D010750:Phosphoproteins; D012269:Ribosomal Proteins; D029701:Saccharomyces cerevisiae Proteins; C031937:phosphoprotein P2, ribosomal; C035690:ribosomal phosphoprotein P1
Biochemistry. 1997 Nov 25;36(47):14439-46
9398162
Mol Microbiol. 2006 Nov;62(4):1191-202
17040491
Science. 1986 Oct 10;234(4773):179-86
3018930
EMBO J. 2000 Nov 15;19(22):6075-84
11080154
Mol Cell Biol. 1995 Sep;15(9):4754-62
7651393
J Comput Chem. 2005 Dec;26(16):1781-802
16222654
Science. 1989 Jun 23;244(4911):1480-3
2544030
J Biol Chem. 2001 Aug 31;276(35):32474-9
11431471
Yeast. 2010 Sep;27(9):693-704
20225338
J Cell Biol. 2009 Sep 21;186(6):849-62
19797078
Proc Natl Acad Sci U S A. 1989 Nov;86(21):8247-51
2682640
Biochim Biophys Acta. 1990 Aug 27;1050(1-3):51-5
2207168
Biochem Cell Biol. 1995 Nov-Dec;73(11-12):959-68
8722011
Int J Biochem Cell Biol. 2000 Jul;32(7):737-46
10856704
Mol Microbiol. 2006 Apr;60(2):386-400
16573688
J Biol Chem. 1997 Feb 7;272(6):3302-8
9013569
J Cell Biol. 2009 Sep 21;186(6):863-80
19797079
Eur J Biochem. 1991 Mar 14;196(2):407-14
1706664
Proc Natl Acad Sci U S A. 2005 Jun 7;102(23):8192-7
15923259
Biochemistry. 2005 Apr 12;44(14):5532-40
15807547
Biochemistry. 1993 Apr 27;32(16):4231-6
8476850
Eur J Biochem. 1979 Aug 1;98(2):409-16
114391
J Biol Chem. 1993 Feb 5;268(4):2451-7
8428920
Proc Natl Acad Sci U S A. 1996 Sep 17;93(19):10315-20
8816797
Int J Biochem Cell Biol. 2002 Aug;34(8):911-5
12007628
J Biol Chem. 1992 Mar 15;267(8):5442-5
1544921
J Biochem. 1985 Dec;98(6):1427-31
4093437
J Biol Chem. 1992 Jun 15;267(17):12061-7
1601875
J Mol Evol. 2008 Aug;67(2):154-67
18612675
J Biol Chem. 2005 Nov 25;280(47):39193-9
16188884
FEBS Lett. 1976 Apr 15;64(1):135-8
773698
J Biol Chem. 1976 Mar 25;251(6):1799-807
767341
J Biol Chem. 1988 Jun 15;263(17):8443-9
3372535
Biol Cell. 2003 May-Jun;95(3-4):179-93
12867082
Eur J Biochem. 1986 Oct 15;160(2):297-304
3769929
J Biol Chem. 1994 Feb 11;269(6):3968-75
8307952
J Biol Chem. 2010 Feb 12;285(7):4747-56
20007716
Nucleic Acids Res. 2010 Aug;38(15):5206-16
20385603
Prog Nucleic Acid Res Mol Biol. 1996;55:157-93
8787610
Mol Cell Biol. 1990 May;10(5):2182-90
2183022
Cell. 2005 Jul 1;121(7):991-1004
15989950
Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10563-7
1961721
Curr Protoc Bioinformatics. 2006 Oct;Chapter 5:Unit-5.6
18428767
Mol Microbiol. 2003 Oct;50(2):703-12
14617190
Int J Biochem Cell Biol. 2009 Jun;41(6):1315-22
19084076
Nucleic Acids Res. 2009 Dec;37(22):7519-32
19789271