Role of a peptide tagging system in degradation of proteins synthesized from damaged messenger RNA

Science

Volumn 271, Issue 5251, 1996, Pages 990-993

  Keiler, Kenneth C ^a   Waller, Patrick R H ^a   Sauer, Robert T ^a  

^a MASSACHUSETTS INSTITUTE OF TECHNOLOGY   (United States)

Author keywords

[No Author keywords available]

Indexed keywords

BACTERIAL ENZYME; CYTOCHROME B; MESSENGER RNA; MUTANT PROTEIN; PEPTIDE; PROTEIN; PROTEINASE; REPRESSOR PROTEIN;

ARTICLE; CARBOXY TERMINAL SEQUENCE; CONTROLLED STUDY; ESCHERICHIA COLI; MOLECULAR RECOGNITION; NONHUMAN; PRIORITY JOURNAL; PROTEIN DEGRADATION; PROTEIN MODIFICATION; STOP CODON; TRANSLATION REGULATION;

EID: 0030024281     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.271.5251.990     Document Type: Article

References (41)

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3. Abbreviations for the amino acid residues are as follows. A, Ala, C, Gys; D, Asp; E, Glu, F, Phe, G, Gly, H, His; I, Ile, K, Lys; L, Leu; M, Met, N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser, T, Thr; V, Val; W, Trp; and Y, Tyr.
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23. 6-trpAt protein may also be degraded by Tsp because its COOH-terminal alanine is the residue most preferred by this protease (K. C. Keiler and R. T. Sauer, J Biol. Chem. 271, 2589 (1996).
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27. - strain. These differ in having five, six, or seven trpA-encoded residues. Similarly, numerous different peptide-tagged forms of IL-6 were identified by Tu et al. (3) We assume that these tagged IL-6 proteins were not degraded because they form inclusion bodies. Size heterogeneity of the IL-6 mRNA was also observed (3). By our model, mRNA heterogeneity naturally results in heterogeneity of the tagged protein products. This mRNA heterogeneity could arise from heterogeneity in transcriptional termination or by nuclease cleavage of full-length mRNA (73) [E Hajnsdorf, O. Steier, L. Coscoy, L. Teysset, P. Regnier, EMBO J. 13, 3368 (1994); H, Causton, B. Py, R. S. McLaren, C. F Higgins, Mol. Microbiol. 14, 731 (1994)].
28. The simplest model is that tagging occurs when an mRNA lacks a translational termination codon. However, the results do not exclude a possible role for the trpAt sequence.
29. - mutants have phenotypes that reflect changes in intracellular proteolysis (26) and alterations in the lysis-lysogeny decisions of temperature bactenophages [D. M. Retallack, L. J. Johnson, D. I. Friedman, J. Bacteriol. 176, 2082 (1994)]. It is unclear whether these phenotypes arise directly or indirectly from loss of the peptide tagging activity of ssrA RNA, with subsequent changes in targeted degradation, or are caused by other defects associated with loss of ssrA RNA [D. M. Retallack and D. I. Friedman, Cell 83, 227 (1995)].
30. - mutants have phenotypes that reflect changes in intracellular proteolysis (26) and alterations in the lysis-lysogeny decisions of temperature bactenophages [D. M. Retallack, L. J. Johnson, D. I. Friedman, J. Bacteriol. 176, 2082 (1994)]. It is unclear whether these phenotypes arise directly or indirectly from loss of the peptide tagging activity of ssrA RNA, with subsequent changes in targeted degradation, or are caused by other defects associated with loss of ssrA RNA [D. M. Retallack and D. I. Friedman, Cell 83, 227 (1995)].
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39. 35S-labeled proteins were detected by autoradiography or analysis with a Phosphorlmager (Molecular Dynamics) All half-lives were calculated from results for a minimum of six time points by nonlinear least-squares fitting of data to an exponential decay. For these calculations, the intensities of the induced bands were integrated with Phosphorimager ImageQuant software and were normalized to a set of stable bands
40. Strain X90 ssrA1::cat was constructed by P1 transduction of the ssrA1::cat allele from E. coli strain JK6257 (26) into E coli strain X90
41. 2-terminal sequencing by sequential Edman degradation; and the Harvard Microchemistry Facility for ion spray mass spectrometry. Supported by NIH grants Al-15706 and Al-16892