Structure of the linkage between adenovirus DNA and the 55,000 molecular weight terminal protein.
Structure of the linkage between adenovirus DNA and the 55,000 molecular weight terminal protein.
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腺病毒 DNA 和 55,000 分子量末端蛋白之间的连接结构。
DOI：
10.1016/0022-2836(81)90208-4
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发表时间：
1981
影响因子：
5.6
通讯作者：
KellyJr,TJ
中科院分区：
文献类型：
--
作者：
Desiderio,SV;KellyJr,TJ
从病毒颗粒中分离出的腺病毒 DNA 的每条互补链的 5' 末端均与表观分子量为 55,000 的蛋白质共价连接。我们已经确定了蛋白质-DNA 连接的结构。 DNase 消化均匀 32P 标记的腺病毒 5 (Ad5) DNA-蛋白质复合物后，分离出与小 [32P] 寡核苷酸连接的 55,000Mr 蛋白质。用胰蛋白酶消化蛋白质并分析所得的[32P]肽，结果如下。 (1)酸水解释放出单个磷酸化氨基酸，在四个独立的电泳或色谱系统中将其鉴定为O-磷酸丝氨酸； (2)用蛇毒磷酸二酯酶处理，如预期仅产生dAMP、dCMP和dTMP(Ad5 DNA 5'端的前25个核苷酸中没有鸟苷酸残基)； (3)预先用蛇毒磷酸二酯酶处理[32P]肽制剂，大大降低了随后酸水解时O-磷酸丝氨酸的产率。这些结果表明 Ad5 DNA 通过丝氨酸残基的 β-OH 上的磷酸二酯键与末端蛋白结合。 DNA-蛋白质连接在碱中很容易水解的发现支持了这一结论。在 70 °C 的 50 mm-NaOH 中，键的水解半衰期约为 10 分钟。在这些条件下孵育Ad5 DNA后，我们能够通过用碱性磷酸酶和多核苷酸激酶连续处理来用32P标记5'末端。将末端标记的 DNA 消化为 5' 单核苷酸，产生 [32P]dCMP。我们得出结论，末端蛋白通过蛋白质丝氨酸残基的 β-OH 和 DNA 末端脱氧胞苷残基的 5'-OH 之间的磷酸二酯键与 Ad5 DNA 结合。
The 5′ terminus of each complementary strand of adenovirus DNA isolated from virions is covalently linked to a protein with an apparent molecular weight of 55,000. We have determined the structure of the protein-DNA linkage. The 55,000Mrprotein, linked to a small [32P]oligonucleotide, was isolated after DNase digestion of uniformly32P-labeled adenovirus 5 (Ad5) DNA-protein complex. The protein was digested with trypsin and the resulting [32P] peptides were analyzed with the following results. (1) Acid hydrolysis released a single phosphorylated amino acid which was identified asO-phosphoserine in four separate electrophoretic or chromatographic systems; (2) treatment with snake venom phosphodiesterase yielded exclusively dAMP, dCMP and dTMP as expected (there are no guanylate residues in the first 25 nucleotides at the 5′ ends of Ad5 DNA); (3) prior treatment of the [32P]peptide preparation with snake venom phosphodiesterase greatly reduced the yield ofO-phosphoserine upon subsequent acid hydrolysis. These results suggest that Ad5 DNA is bound to the terminal protein by a phosphodiester linkage to the β-OH of a serine residue. This conclusion is supported by the finding that the DNA-protein linkage is readily hydrolyzed in alkali. In 50 mm-NaOH at 70 °C the half time for hydrolysis of the linkage is about ten minutes. After incubation of Ad5 DNA under these conditions we were able to label the 5′ termini with32P by sequential treatment with alkaline phosphatase and polynucleotide kinase. Digestion of the end-labeled DNA to 5′ mononucleotides yielded [32P]dCMP. We conclude that the terminal protein is bound to Ad5 DNA by a phosphodiester linkage between the β-OH of a serine residue of the protein and the 5′-OH of the terminal deoxycytidine residue of the DNA.
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影响因子：
64.5
作者：
REKOSH, DMK;RUSSELL, WC;ROBINSON, AJ
通讯作者：
ROBINSON, AJ
影响因子：
5.6
作者：
KELLY, TJ;SMITH, HO
通讯作者：
SMITH, HO
影响因子：
5.6
作者：
CHALLBERG, MD;KELLY, TJ
通讯作者：
KELLY, TJ
转移 RNA 中的次要成分：它们的特征、位置和功能。
DOI：
--
发表时间：
1972
期刊：
影响因子：
--
作者：
Susumu Nishimuka
通讯作者：
Susumu Nishimuka
人类腺病毒 DNA 5 末端被封闭的证据。
影响因子：
3.7
作者：
E. Carusi
通讯作者：
E. Carusi