The N‐Terminal Sequence Protein of L7/L12 Is Responsible for Its Dimerization

Gudkov, A.T.; Behlke, Joachim

doi:10.1111/j.1432-1033.1978.tb12605.x

Cited by 65 publications

(60 citation statements)

References 10 publications

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“…Indeed, very recent evidence strongly suggests that the CTD of L7͞L12 is responsible for elongation-factor Tu (EF-Tu) function (37). The NTD (residues 1-36) of L7͞L12 mediates the dimer interaction and anchors the protein, via L10, to the slow-tumbling ribosomal body (38). In accord with this conclusion, no resonances of the NTD domain can be observed in the NMR spectra.…”

Section: Discussionmentioning

confidence: 99%

Heteronuclear NMR investigations of dynamic regions of intact Escherichia coli ribosomes

Christodoulou

Larsson

Fucini

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

121

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15 N-1 H NMR spectroscopy has been used to probe the dynamic properties of uniformly 15 N labeled Escherichia coli ribosomes. Despite the high molecular weight of the complex (Ϸ2.3 MDa), [ 1 H-15 N] heteronuclear single-quantum correlation spectra contain Ϸ100 well resolved resonances, the majority of which arise from two of the four C-terminal domains of the stalk proteins, L7͞L12. Heteronuclear pulse-field gradient NMR experiments show that the resonances arise from species with a translational diffusion constant consistent with that of the intact ribosome. Longitudinal relaxation time (T1) and T1 15 N-spin relaxation measurements show that the observable domains tumble anisotropically, with an apparent rotational correlation time significantly longer than that expected for a free L7͞L12 domain but much shorter than expected for a protein rigidly incorporated within the ribosomal particle. The relaxation data allow the ribosomally bound C-terminal domains to be oriented relative to the rotational diffusion tensor. Binding of elongation factor G to the ribosome results in the disappearance of the resonances of the L7͞L12 domains, indicating a dramatic reduction in their mobility. This result is in agreement with cryoelectron microscopy studies showing that the ribosomal stalk assumes a single rigid orientation upon elongation factor G binding. As well as providing information about the dynamical properties of L7͞L12, these results demonstrate the utility of heteronuclear NMR in the study of mobile regions of large biological complexes and form the basis for further NMR studies of functional ribosomal complexes in the context of protein synthesis. P rotein synthesis in living systems takes place on the ribosome, a complex macromolecular assembly whose structural and functional properties are rapidly emerging from a powerful combination of electron microscopy (EM) and x-ray crystallography (1, 2). In Escherichia coli, the ribosome is composed of 54 different proteins and three RNA molecules (23S, 16S, and 5S rRNA) This 2.3-MDa complex is termed the 70S ribosome and is made up of two components, the 30S and 50S subunits. The translation of genetic information into functional proteins involves a number of auxiliary factors, many of which are GTPases, including IF2, EF-Tu, elongation factor G (EF-G), and RF3 (2). These molecules bind to overlapping sites on the 50S subunit and regulate the transition of the ribosome through various states on the translational pathway. The binding sites are collectively known as the GTPase-associated region, due to the role of this region in stimulating the GTPase activity of the auxiliary factors.The GTPase-associated region (GAR) includes helices 42-44 and 95 (the ␣-sarcin loop) of 23S RNA, and the proteins L10, L11, and L7͞L12 (1). The latter (L7͞L12) is located on the ribosomal stalk and is unique among the ribosomal proteins, because it is the only protein present in multiple copies (four proteins per ribosomal particle). Although atomic details of much of the GAR have been reve...

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Section: Discussionmentioning

confidence: 99%

Heteronuclear NMR investigations of dynamic regions of intact Escherichia coli ribosomes

Christodoulou

Larsson

Fucini

et al. 2004

Proc. Natl. Acad. Sci. U.S.A.

121

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“…Considering the data cited above, we can conclude that formation of the S-S bond in the mutant protein does not influence the flexibility of the hinge region (residues 38-50), which is functionally important [18,19] and does not distort the structure of the N-terminal sequence which is responsible for protein dimerization [4,6].…”

Section: Resultsmentioning

confidence: 99%

“…Thus, the N-terminal part of the protein is important for its dimerization and activity [4,6,7]. According to NMR studies the C-terminal globules have a high mobility in the 70S ribosomes [8].…”

Section: Introductionmentioning

confidence: 99%

The first 37 residues are sufficient for dimerization of ribosomal L7/L12 protein

1995

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“…Several lines of evidence indicate that the N-terminal domains of P1/P2, and bacterial equivalents are involved in dimerisation (Gonzalo et al, 2001;Gudkov and Behlke, 1978;Kopke et al, 1992;Tchorzewski et al, 2000; and in interaction with P0 or L10 (Gonzalo et al, 2001;Kopke et al, 1992;Pettersson et al, 1976;Zurdo et al, 2000).…”

Section: Elements Involved In the Dimerisation Of P1/p2 (And Equivalementioning

confidence: 99%

The puzzling lateral flexible stalk of the ribosome

Gonzalo

Reboud

2003

Biology of the Cell

127

131

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The lateral flexible stalk of the large ribosomal subunit is made of several interacting proteins anchored to a conserved region of the 28S (26S) rRNA termed the GTPase-associated domain or thiostrepton loop. This structure is demonstrated to adopt puzzling changes of conformation following the different steps of the elongation cycle. Some of these proteins termed the P-proteins in eukaryotes and L10 and L7/L12 in bacteria, present little structural similarities between Eubacteria on one side and Archae and Eukaryotes on the other side. However, up to now, these proteins seem to present a similar macromolecular organisation and they have been involved in the same functions. Convincing evidence attests that these proteins participate in elongation factor binding to the ribosome, and it has been suggested that these proteins might be evolved in a GTP hydrolysis activating protein activity. Involvement of these proteins in the translational mechanism is discussed. Moreover, in eukaryotes, small P-proteins are also found as isolated proteins in a cytoplasmic pool that exchanges with the ribosome-associated P-proteins. Moreover, a part of the ribosomal proteins is phosphorylated (hence their P-protein names). The biological signification of these particularities is discussed.

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The N‐Terminal Sequence Protein of L7/L12 Is Responsible for Its Dimerization

Cited by 65 publications

References 10 publications

Heteronuclear NMR investigations of dynamic regions of intact Escherichia coli ribosomes

Heteronuclear NMR investigations of dynamic regions of intact Escherichia coli ribosomes

The first 37 residues are sufficient for dimerization of ribosomal L7/L12 protein

The puzzling lateral flexible stalk of the ribosome

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