Abstract
By a combination of protein chemistry and recombinant DNA methods a glycine-rich region was found to be located near the carboxyl terminus of the nucleolar specific phosphoprotein, nucleolin, from Novikoff hepatoma (protein C23) and Chinese hamster ovary cells (100-kDa nucleolar protein). A sequence of 192 amino acid residues was derived from partial sequences of cyanogen bromide and N-bromosuccinimide fragments of protein C23 and deduced protein sequence from Chinese hamster ovary cell 100-kDa cDNA sequences. The 66 residues sequenced by protein methods were identical to the corresponding residues deduced by DNA sequencing. The multiple residues of NG,NG-dimethylarginine (DMA) contained in the nucleolin polypeptide were found to be limited to a segment of less than 10 kDa near the carboxyl-terminal end of the protein. This segment also contained internally repeated sequences (e.g. 7 copies of the sequence Gly-Gly-Arg-Gly-Gly were found) which were unrelated to sequences closer to the amino-terminal end. Most arginine residues in this region were surrounded by 2 or 3 glycine residues and were relatively close in sequence to phenylalanine residues.
Highlights
From the Centre deRecherche de Bwchemie et de Genetique Cellulaires,Centre Natwnul de la RechercheScientifique, 118, route de Narbonne, 31062 Toulouse Cedex, France
To be located near the carboxyl termionfutshe nucleo- little is known about how the nulceolar subcomlar specific phosphoprotein, nucleolin, from Novikoff ponents are organized or how the process of ribosome biohepatoma(protein C23) andChinesehamsterovary genesis takes place, recent work in various laboratories has cells (100-kDa nucleolar protein)
NG,p-dimethylarginine(DMA) contained in the nu- analogous proteins have been found in other systems such as cleolin polypeptide were found to be limitedto a seg- mouse ascites sarcoma cells by Kawashima et a1. [8]and in various mouse, hamster, end of the protein
Summary
From the Centre deRecherche de Bwchemie et de Genetique Cellulaires,Centre Natwnul de la RechercheScientifique, 118, route de Narbonne, 31062 Toulouse Cedex, France. The multiple residueosf having molecular masses of 100 to 110 kDa. Apparently NG,p-dimethylarginine(DMA) contained in the nu- analogous proteins have been found in other systems such as cleolin polypeptide were found to be limitedto a seg- mouse ascites sarcoma cells (nucleolar specific phosphoproment of less than 10 kDa near the carboxyl-terminal tein) by Kawashima et a1. NG,p-dimethylarginine(DMA) contained in the nu- analogous proteins have been found in other systems such as cleolin polypeptide were found to be limitedto a seg- mouse ascites sarcoma cells (nucleolar specific phosphoproment of less than 10 kDa near the carboxyl-terminal tein) by Kawashima et a1. [8]and in various mouse, hamster, end of the protein This segment contained inter- human, Drosophila, and chicken cell lines (pp105) by Pfeifle nally repeated sequences(e.g. 7 copies of the sequence and Anderer [10].In virtually all of these studies, polypeptides. Nature andabundance in nucleoli of this polypeptide we will refer to it as “nucleolin” and specify the species of origin where appropriate
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