Summary
Deletions in the promoter region of the 37S pre-rRNA operon in yeast were constructed and analysed in vivo using an artificial ribosomal minigene present on an extrachromosomal yeast vector. Sequences required for correct transcription initiation were found to be located between positions −192 and +15 relative to the start; a 5′-deletion down to position −133 reduces the transcription yield of the minigene at least five-fold. To allow detection of transcription of the minigene in isolated nuclei of yeast transformed with a minigene-bearing plasmid we attempted to increase the minigene copy number. The transcription yield in vivo appeared not to be proportional to the copy number but was found to be greatly enhanced when two or three mini-genes are present in tandem. α-Amanitin sensitivity of transcription of these minigenes in isolated nuclei proved that RNA polymerase I is responsible for their transcription.
Similar content being viewed by others
References
Beggs JD (1981) Alfred Benzon Symp 16:383
Breuning KD, Macedonski V, Hollenberg CP (1982) Gene 20:1–10
Broach J (1983) Methods Enzymol 101:307–325
Dierkes P, van Ooyen A, Cochran MD, Dobkin C, Reisner J, Weissmann C (1983) Cell 32:695–706
Elion EA, Warner JR (1984) Cell 39:663–673
Erhart E, Hollenberg CP (1983) J Bacteriol 156:625–635
Grummt I (1982) Proc Natl Acad Sci USA 79:6908–6911
Grummt I, Roth E, Paule M (1982) Nature (London) 296:173–174
Grummt I, Skinner JA (1985) Proc Natl Acad Sci USA 82:722–726
Hollenberg CP (1979) Extrachromosomal DNA. In: Cummings D, Borst P, Dawid I, Weissman S, Fox CF (eds) ICN-UCLA Symp, vol 15. Academic Press, New York, pp 325–338
Ide GI, Saunders CA (1981) Curr Genet 4:85–90
Kempers-Veenstra AE, van Heerikhuizen H, Musters W, Klootwijk J, Planta RJ (1984) EMBO J 3:1377–1382
Klootwijk J, Verbeet MPh, Veldman GM, de Regt VCHF, van Heerikhuizen H, Planta RJ (1984) Nucleic Acids Res 12:1377–1390
Kohorn BD, Rae PMM (1983) Proc Natl Acad Sci USA 80:3265–3268
Learned RM, Smale ST, Haltiner MM, Tjian R (1983) Proc Natl Acad Sci USA 80:3558–3562
Lohr D, Ide GI (1983) J Biol Chem 258:4668–4671
Long OE, Dawid IB (1980) Annu Rev Biochem 49:727–764
Maxam AM, Gilbert W (1977) Proc Natl Acad Sci USA 74:560–564
Meyerink JH (1979) PhD thesis, Vrije Universiteit Amsterdam
Miller KG, Tower J, Sollner-Webb B (1985) Mol Cell Biol 5:554–562
Molenaar CMTh (1984) PhD thesis, Vrije Universiteit Amsterdam
Moss T (1982) Cell 30:835–842
Moss T (1983) Nature (London) 302:223–228
Nomura M, Gourse R, Baughman G (1984) Annu Rev Biochem 53:75–117
Paule M, Iida CT, Perna PJ, Harris GH, Knoll PA, D'Allessio JM (1984) Nucleic Acids Res 12:8161–8180
Pedersen MB (1983) Carlsberg Res Commun 48:485–500
Reeder RH, Wilkinson J, Bakken A, Morgan G, Busby SJ, Roan J, Sollner-Webb B (1982) Cold Spring Harbor Symp Quant Biol 57:867–871
Reeder RH (1984) Cell 38:349–351
Sarmientos P, Cashel M (1983) Proc Natl Acad Sci USA 80:7010–7013
Schultz LD, Hall BD (1976) Proc Natl Acad Sci USA 73:1029–1033
Skinner JA, Öhrlein A, Grummt I (1984) Proc Natl Acad Sci USA 81:2137–2141
Sollner-Webb B, Wilkinson J, Roan J, Reeder RH (1983) Cell 35:199–206
Southern EM (1975) J Mol Biol 98:503–517
Valenzuela P, Hager GL, Weinberg F, Rutter WJ (1976) Proc Natl Acad Sci USA 73:1024–1028
Verbeet MPh, Klootwijk J, van Heerikhuizen H, Fontijn RD, Vreugdenhil E, Planta RJ (1984) Nucleic Acids Res 12:1137–1148
Yamamoto I, Takakusa N, Mishima Y, Kominami R, Muramatsu M (1984) Proc Natl Acad Sci USA 81:299–303
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Kempers-Veenstra, A.E., Musters, W., Dekker, A.F. et al. Deletion mapping of the yeast pol I promoter. Curr Genet 10, 253–260 (1985). https://doi.org/10.1007/BF00365621
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00365621