Devine J M, Tsang A S, Williams J G
Cell. 1982 Apr;28(4):793-800. doi: 10.1016/0092-8674(82)90058-7.
Discoidin I and II are lectins synthesized during the aggregation of Dictyostelium discoideum amoebae which may play a role in cellular cohesion. Discoidin I was thought to consist of two major polypeptides, but we show that there are three. The N-terminal amino acid sequence of the polypeptides has been predicted by determining part of the nucleotide sequence of their respective mRNAs. We obtained the nucleotide sequences by reverse transcription of the mRNAs, using as primers, fragments derived from the coding region of two cloned discoidin I sequences, and utilizing cross hybridization to the various mRNA species and differences in the length of their 5' noncoding regions to isolate fragments for DNA sequencing. We used primer extension to measure the relative concentration of the three major discoidin I mRNA sequences. We show that during development changes in the abundance of all three mRNA sequences occur coordinately. In cells growing in nutrient broth, however, only two of the three major discoidin I mRNA sequences accumulate, and if such cells are grown to a very high density, both sequences disappear. These results indicate that the coordination of discoidin I gene expression is not obligatory and that the members of this multigene family may differ in the mode of their induction during normal development.
盘状蛋白I和II是在盘基网柄菌变形虫聚集过程中合成的凝集素,可能在细胞黏附中发挥作用。盘状蛋白I曾被认为由两种主要多肽组成,但我们发现有三种。通过确定其各自mRNA的部分核苷酸序列,预测了这些多肽的N端氨基酸序列。我们通过mRNA的逆转录获得核苷酸序列,使用来自两个克隆的盘状蛋白I序列编码区的片段作为引物,并利用与各种mRNA种类的交叉杂交以及它们5'非编码区长度的差异来分离用于DNA测序的片段。我们使用引物延伸来测量三种主要盘状蛋白I mRNA序列的相对浓度。我们表明,在发育过程中,所有三种mRNA序列丰度的变化是协同发生的。然而,在营养肉汤中生长的细胞中,三种主要盘状蛋白I mRNA序列中只有两种积累,如果将这些细胞培养到非常高的密度,这两种序列都会消失。这些结果表明,盘状蛋白I基因表达的协调并非必然,并且这个多基因家族的成员在正常发育过程中的诱导方式可能不同。