In order to study the biosynthesis of ribosomal RNA in Saccharomyces carlsbergensis the labelling kinetics of the various precursor and mature rRNA species were determined using pulse-labelling of protoplasts with [5-3H] uridine at 15 degrees C. Label appears almost immediately in 37 S RNA, the precursor common to both 26 S and 17 S rRNA. Labelled 29 S and 18 S RNA, the immediate precursors of 26 S and 17 S rRNA respectively, were found to appear about 4 min and about 8 min after addition of the isotope respectively. These data indicate that the topography of the 37 S precursor RNA is: 5'-17 S -26 S-3'. The pool size of 29 S RNA is about twice as large as that of either 37 S or 18 S RNA, indicating that under the conditions used processing of 18 S to 17 S rRNA proceeds more rapidly than processing of 29 S to 26 S rRNA. The labelling kinetics of 5.8 S rRNA are in agreement with the existence of a 7 S precursor rRNA, the identity of which was previously established (Trapman, J., de Jonge, P. and Planta, R.J. (1975) FEBS Lett. 57, 26--30) and which, in turn, probably is derived from 29 S precursor rRNA. The labelling kinetics of 5 S rRNA suggest that 5 S RNA sequences, rather than also being part of the common 37 S precursor, are located on a separate primary transcription product. Whether this transcript still contains excess sequences remains to be determined. However, because of the rapid appearance of labelled 5 S RNA, such a precursor would have to be very short lived.