Skip to main content
SpringerLink
Log in

The peptidyl-puromycin reaction with Neurospora crassa ribosomes

The number of active ribosomes in different conditions of growth

  • Published:
Archiv für Mikrobiologie Aims and scope Submit manuscript

Summary

The reaction of the peptidyl-tRNA in an “in vitro” system from Neurospora crassa with puromycin has been studied by different experimental approaches. Ribosomes precharged with labelled polyphenylalanine have been prepared and the release of radioactivity, occurring after the reaction with puromycin, has been followed on sucrose density gradients. Furthermore the reaction of endogenous peptidyl-tRNA carried by ribosomes isolated from actively growing mycelia with 3H-puromycin has been characterized. By this latter technique it has been possible to evaluate the percentage of ribosomes engaged in protein synthesis in ribosomal populations isolated from mycelia in different stages of growth.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

TF-1:

aminoacyl transferase I

TF-2:

aminoacyl transferase II

PM:

puromycin

3H-PM:

puromycin-methoxy-3H dihydrochloride

HEPES:

N-2-Hydroxyethylpiperazine-N′-2-ethanesulfonic acid

TCA:

trichloroacetic acid

PEP:

phosphoenolpyruvate

tRNA:

transfer RNA

rRNA:

ribosomal RNA

PPO:

2,5-diphenyloxazole

POPOP:

1,4-bis-(5-phenyloxazol-2-yl) benzene

References

  • Alberghina, F. A. M., Chimenti Signorini, R., Trezzi, F., Viotti, A. Effect of magnesium on growth and morphology of Neurospora crassa. J. Submicr. Cytol. 3, 9–18 (1971).

    Google Scholar 

  • —, Sassella, D., Sturani, E.: Ribosomes of Neurospora crassa: changes in the magnesium dependence of the poly U directed phenylalanine polymerization. G. Bot. Ital. 104, 249–262 (1970).

    Google Scholar 

  • —, Sturani, E.: RNA content and rate of growth in Neurospora crassa mycelia. Neurospora Newsl. 18, 9–10 (1971).

    Google Scholar 

  • ——, Ursino, D. J., Marrè, E.: Ribosomal changes related to changes in the rate of growth in Neurospora crassa mycelia. Biochim. biophys. Acta (Amst.) 195, 576–578 (1969).

    Google Scholar 

  • Click, R. E., Tint, B. L.: Comparative sedimentation rates of plant, bacterial and animal ribosomal RNA. J. molec. Biol. 25, 111–122 (1967).

    Google Scholar 

  • Dure, L. S., Epler, J. L., Barnett, B. E.: Sedimentation properties of mitochondrial and cytoplasmic ribosomal RNAs from Neurospora. Proc. nat. Acad. Sci. (Wash.) 58, 1883–1887 (1967).

    Google Scholar 

  • Gonzales, N. S., Goldenberg S. H., Algranati, I. D.: Protein synthesis and ribosomal distribution at different growth stages in Bacillus stearothermophilus. Biochim. biophys. Acta (Amst.) 168, 760–762 (1968).

    Google Scholar 

  • Kjeldgaard, N. O.: Regulation of nucleic acid and protein formation in bacteria. Advanc. Microbiol. Phys. 1, 39–95 (1967).

    Google Scholar 

  • Lengyel, P.: The process of translation as seen in 1969. Cold Spr. Harb. Symp. quant. Biol. 34, 828–841 (1969).

    Google Scholar 

  • Maaloe, O., Kjeldgaard, N. O.: Control of macromolecular synthesis, pp. 70–124. New York-Amsterdam: W. A. Benjamin 1966.

    Google Scholar 

  • Phillips, L. A., Franklin, R. M.: The in vivo distribution of bacterial polysomes, ribosomes and ribosomal subunits. Cold Spr. Harb. Symp. quant. Biol. 34, 243–253 (1969).

    Google Scholar 

  • Sistrom, W. R.: Microbial life, p. 108, Second edit. New York: Holt, Rinehart and Winstin Inc. 1969.

    Google Scholar 

  • Stanners, C. P., Becker, H.: Control of macromolecular synthesis in proliferating and resting Syrian hamster cells in monolayer culture: I. Ribosome function. J. Cell Physiol. 77, 31–42 (1971).

    Google Scholar 

  • Sturani, E., Alberghina, F. A. M.: A poly U directed phenylalanine polymerizing system from Neurospora crassa: General characterization and ionic effects. Ital. J. Biochem. 19, 319–336 (1970).

    Google Scholar 

  • Subramanian, A. R., Davis, B. D., Beller, R. J.: The ribosome dissociation factor and the ribosome-polysome cycle. Cold Spr. Harb. Symp. quant. Biol. 34, 223–230 (1969).

    Google Scholar 

  • Tiboni, O., Parisi, B., Perani, A., Ciferri, O.: Presence of two sets of ribosome specific transfer factors in the cell-free extracts from non-photosynthetic alga Prototheca zopfii. J. molec. Biol. 47, 467–476 (1970).

    Google Scholar 

  • Viotti, A., Bagni, N., Sturani, E., Alberghina, F. A. M.: Magnesium and polyamines levels in Neurospora crassa mycelia. Biochim. biophys. Acta (Amst.) 244, 329–337 (1971).

    Google Scholar 

  • Wool, I. G., Kurihara, K.: Determination of the number of active muscle ribosomes: effect of diabetes and insulin. Proc. nat. Acad. Sci. (Wash.) 58, 2401–2407 (1967).

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Laboratorio di Fisiologia Vegetale, Istituto di Scienze Botaniche della Università di Milano, Centro del C.N.R. di Biologia Molecolare e Cellulare delle piante, Milano, Italy

    F. A. M. Alberghina, E. Sturani & L. Schiaffonati

  2. Institute of Plant Sciences, University of Milan, Via G. Colombo, 60, I-20133, Milan, Italy

    F. A. M. Alberghina

Authors
  1. F. A. M. Alberghina
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. E. Sturani
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. L. Schiaffonati
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Alberghina, F.A.M., Sturani, E. & Schiaffonati, L. The peptidyl-puromycin reaction with Neurospora crassa ribosomes. Archiv. Mikrobiol. 80, 166–175 (1971). https://doi.org/10.1007/BF00411881

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00411881

Keywords

Search

Navigation