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R L Chisholm

Publications and source records attributed to R L Chisholm.

41 records · Page 3Linked to original sources

A repetitive Dictyostelium gene family that is induced during differentiation and by heat shock.

Clone pB41-6 (2.5 kb) contains sequences that are repeated 200-300 times in the Dictyostelium genome; about 40 of these sequences are part of a 4.5 kb repeated and apparently transposable genomic element. Clone pB41-6 hybridizes to a large number of cytoplasmic polyadenylated RNAs whose accumulation begins in the first hour of differentiation. In order to understand the regulation of these repeated sequences, we have sequenced pB41-6. It contains three long open reading frames in the "sense" strand. Remarkably, about 70 bases upstream of the transcription initiation site is a sequence identical to that responsible for induction of the Drosophila heat shock genes. A search of published sequences also generated a similar sequence upstream of one of the Dictyostelium actin genes. Indeed, we found that both pB41-6-related RNAs and actin mRNAs are increased as a result of heat shocking growing cells, and that transcription of pB41-6 sequences is induced by heat shock. Thus Dictyostelium contains a set of genes that are induced as a response to heat shock or to the stresses that trigger the initiation of development. We show here that the principal component of this "stress" is not amino acid starvation but the high density of the cells.

Actins↗

Different mRNAs have different nuclear transit times in Dictyostelium discoideum aggregates.

Nuclear processing of mRNA precursors in differentiating multicellular Dictyostelium discoideum aggregates is markedly slower than in growing amoebae. Thus, we have been able to determine the time of nuclear processing of individual mRNA species in postaggregating cells by following the incorporation of 32PO4 into nuclear and cytoplasmic RNA complementary to cloned cDNAs. Precursors of mRNAs synthesized during both growth and differentiation remain in the nucleus for about 25 to 60 min. By contrast, typical mRNAs which are synthesized only by postaggregative cells have nuclear processing times between 50 and 100 min. Depending on the particular mRNA, between 20 and 60% of nuclear transcripts are converted into cytoplasmic mRNA. A third class of mRNAs are transcribed from a set of repetitive DNA segments and are expressed predominantly during differentiation. Nuclear precursors of these mRNAs are extensively degraded within the nucleus or very rapidly after transport to the cytoplasm. Those sequences that are stable in the cytoplasm exit from the nucleus only after a lag of over 2 h. Thus, mRNAs encoded by different genes that are subject to different types of developmental controls display different times of transit to the cytoplasm and different efficiencies of nuclear processing. Differential nuclear processing may contribute to the regulation of the level of individual cytoplasmic mRNAs.

Biological Transport↗

The significance of varying SRBC/lymphocyte ratio in T cell rosette formation.

The incubation ratio of sheep red blood cells (SRBC) to lymphocytes is a critical factor in rosette formation, whereas the length of time SRBC and lymphocytes are incubated together does not significantly affect the percentage of lymphocytes forming rosettes. The graph obtained by plotting percentage of rosette formation against the ratio of SRBC to lymphocytes is similar to that resulting from the formation of bimolecular complexes. If rosette formation is analogous to formation of bimolecular complexes, maximal rosette formation occurs when the system is saturated, i.e., with excess SRBC, and is a measure of the total capacity of a lymphocyte population to form rosettes. In addition, the percentage of rosette formation observed at a limiting SRBC/lymphocyte ratio gives an indication of the avidity of the lymphocytes for SRBC. This interpretation may provide an explanation for the difference between the "active" and "total" rosettes. When the log of the SRBC/lymphocyte ratio is plotted against percentage of rosette formation, a straight line is obtained, suggesting that within a given normal lymphocyte sample, T cell subsets with different avidities are not detected by rosette formation at different SRBC/lymphocyte ratios.

Blood Cell Count↗

Mechanism of sequential induction of cell-type specific mRNAs in Dictyostelium differentiation.

Upon starvation, the cellular slime mould Dictyostelium discoideum initiates a 24-h programme of differentiation. Within 6 h, cells move towards aggregation centres in response to pulsatile synthesis and secretion of cyclic AMP. At about 12 h, aggregates of 10(5) cells are formed, held together by newly made surface adhesion molecules. The cells then differentiate into the two principal types found in the terminal stage of development, spores and stalks. Here we show that the chemotaxis and aggregation stages of this developmental programme can be described as a series of sequential events in which these extracellular signals--starvation, cyclic AMP and cell-cell contact--induce specific, sequential changes in the pattern of gene expression.

Cell Differentiation↗