Hong J C, Nagao R T, Key J L
J Biol Chem. 1987 Jun 15;262(17):8367-76.
A cDNA clone, pTU04, which hybridizes to two different sizes of mRNA on Northern blots was isolated from soybean suspension culture cell poly(A) RNA. Northern analysis reveals that meristematic tissue produces a 1050-nucleotide mRNA while quiescent mature cells produce primarily a 1220-nucleotide mRNA homologous to pTU04. The cDNA and its corresponding genomic clone have been partially characterized. The nucleotide sequence of the gene predicts a proline-rich protein, designated SbPRP1, which contains a signal peptide sequence and 43 repeats of a sequence consisting primarily of Pro-Pro-Val-Tyr-Lys (CCA-CCA-GTT-TAC-AAG). From nuclease S1 and hybrid-select translation analyses, the cDNA clone pTU04 appears to represent the mRNA for the mature tissue 1220-nucleotide RNA observed on Northern blots. Although there is no direct proof that the encoded protein is a cell wall protein, it has the properties similar to previously isolated cell wall proteins: 1) it is very basic with a high content of Pro, Tyr, and Lys; 2) it has similar hydropathic properties; and 3) its repeating unit shares sequence homology with that of more highly characterized cell wall proteins, generally termed extensin (Chen, J., and Varner, J. E. (1985) EMBO J. 4, 2145-2151; Smith, J. J., Muldoon, E. P., Willard, J. J., and Lamport, D. T. A. (1986) Phytochemistry 25, 1021-1030.
从大豆悬浮培养细胞的聚腺苷酸RNA中分离出一个cDNA克隆pTU04,它在Northern杂交印迹上能与两种不同大小的mRNA杂交。Northern分析表明,分生组织产生一种1050个核苷酸的mRNA,而静止的成熟细胞主要产生一种与pTU04同源的1220个核苷酸的mRNA。该cDNA及其相应的基因组克隆已得到部分特征分析。该基因的核苷酸序列预测出一种富含脯氨酸的蛋白质,命名为SbPRP1,它含有一个信号肽序列以及43个主要由脯氨酸-脯氨酸-缬氨酸-酪氨酸-赖氨酸(CCA-CCA-GTT-TAC-AAG)组成的序列重复。通过核酸酶S1和杂交选择翻译分析,cDNA克隆pTU04似乎代表了在Northern杂交印迹上观察到的成熟组织1220个核苷酸RNA的mRNA。尽管没有直接证据表明所编码的蛋白质是细胞壁蛋白,但它具有与先前分离的细胞壁蛋白相似的特性:1)它呈强碱性,脯氨酸、酪氨酸和赖氨酸含量高;2)它具有相似的亲水性;3)其重复单元与特征更明显的细胞壁蛋白(通常称为伸展蛋白)的重复单元具有序列同源性(陈,J.,和瓦尔纳,J.E.(1985年)《欧洲分子生物学组织杂志》4,2145 - 2151;史密斯,J.J.,马尔登,E.P.,威拉德,J.J.,和兰波特,D.T.A.(1986年)《植物化学》25,1021 - 1030)。