Yeh H Y, Pieniazek N, Pieniazek D, Gelderblom H, Luftig R B
Department of Microbiology, Immunology and Parasitology, Louisiana State University Medical Center, New Orleans 70112-1393.
Virus Res. 1994 Aug;33(2):179-98. doi: 10.1016/0168-1702(94)90054-x.
DNA sequencing of the subgroup F human adenovirus serotype 41 (TAK, Ad41) fiber gene revealed the presence of two adjacent open reading frames encoding information for proteins with molecular weights of 60.6 kDa and 41.4 kDa (Pieniazek, et al; Nucleic Acids Res. 18: p. 1901, 1990). In this paper, various approaches were used to characterize the two proteins and determine whether both fibers were expressed in infected cells as well as on viral particles. We initially used a reverse transcriptase-polymerase chain reaction with primers for the short and long fiber genes to amplify mRNA from Ad41 infected HEp-2 cells at 48 h post-infection. Two distinct DNA bands; one slightly larger than 1.1 kbp and the other at about 1.7 kbp were identified. Second, we used polyclonal anti-Ad41 virion and monoclonal anti-Ad5 fiber antibodies to demonstrate that at both 24 and 36 h post-infection, Ad41 expressed two fiber proteins of the expected size. Specifically, by SDS-PAGE, one fiber (short) had a molecular weight of 40 kDa, while the other (long) had a molecular weight of 60 kDa. Third, by electron microscopy, two sizes of fibers were released from CsCl purified virions, both having a characteristic adenovirus morphology, with a knob at one end. The long fiber measured 315A in length and the short fiber was 250A long. These measurements are consistent with the two Ad41 fibers being encoded by the above open reading frames. We also performed a computer search to compare fiber sequences from other human adenovirus serotypes with that of the Ad41 short and long fiber proteins. The primary structure of both Ad41 fibers were found to be similar in that they contained tail, shaft and knob regions. Further, the tail region of both fibers (amino acids 1-42) showed a 74% overall homology to each other and contained the Ad conserved sequence NH2-F-N-P-V-Y-P-Y-COOH. An interesting difference, however, was observed in the shaft region where the long fiber (amino acids 43-389) had twenty-two 16-amino acid repeat motifs, while the short fiber (amino acids 43-233) had only twelve. Finally, we noted that the long fiber knob region was about 15% longer than that of the short fiber, and showed little overall homology. In conclusion, human adenovirus subgroup F (type 41) virions appear to differ from those of all other human adenoviruses (subgenera A-E) in that they contain two fiber genes and correspondingly, two different sized fibers.(ABSTRACT TRUNCATED AT 400 WORDS)
F亚组人类腺病毒41型（TAK，Ad41）纤维基因的DNA测序显示存在两个相邻的开放阅读框，它们编码分子量分别为60.6 kDa和41.4 kDa的蛋白质信息（皮涅亚泽克等人；《核酸研究》18：第1901页，1990年）。在本文中，我们采用了多种方法来表征这两种蛋白质，并确定两种纤维是否在感染细胞以及病毒颗粒上都有表达。我们最初使用针对短纤维基因和长纤维基因的引物进行逆转录聚合酶链反应，以扩增感染后48小时的Ad41感染的HEp - 2细胞中的mRNA。鉴定出两条不同的DNA条带；一条略大于1.1 kbp，另一条约为1.7 kbp。其次，我们使用多克隆抗Ad41病毒体抗体和单克隆抗Ad5纤维抗体来证明，在感染后24小时和36小时，Ad41都表达了预期大小的两种纤维蛋白。具体而言，通过SDS - PAGE分析，一种纤维（短纤维）的分子量为40 kDa，而另一种（长纤维）的分子量为60 kDa。第三，通过电子显微镜观察，从氯化铯纯化的病毒体中释放出两种大小的纤维，两者都具有典型的腺病毒形态，一端有一个球状结构。长纤维长度为315埃，短纤维长度为250埃。这些测量结果与上述开放阅读框编码的两种Ad41纤维一致。我们还进行了计算机搜索，以比较其他人类腺病毒血清型的纤维序列与Ad41短纤维和长纤维蛋白的序列。发现Ad41两种纤维的一级结构相似，都包含尾部、杆部和球状结构区域。此外，两种纤维的尾部区域（氨基酸1 - 42）彼此之间总体同源性为74%，并包含腺病毒保守序列NH2 - F - N - P - V - Y - P - Y - COOH。然而，在杆部区域观察到一个有趣的差异，长纤维（氨基酸43 - 389）有二十二个16氨基酸重复基序，而短纤维（氨基酸43 - 233）只有十二个。最后，我们注意到长纤维的球状结构区域比短纤维长约15%，且总体同源性较低。总之，F亚组人类腺病毒（41型）病毒体似乎与所有其他人类腺病毒（A - E亚属）不同，因为它们含有两个纤维基因，相应地，有两种不同大小的纤维。（摘要截短至400字）
