New human gene that mediates herpes simplex virus entry

描述(由申请人提供)：这项研究启动了对一种新分离的人类基因的研究，目前命名为人胎肺cDNAA27(HFI-A27)。它是从人胎肺cDNA文库的功能筛选中分离到的三个新的人类基因之一。我们寻找将HSV进入易感性转移到缺乏HSV进入受体的猪细胞的基因。HFL-A27的核苷酸序列含有一个开放阅读框，可产生37kD和28kD的体外翻译蛋白。其基因产物在猪细胞中瞬时表达时可介导HSV-1进入。目标是开发试剂并获得关于HFL-A27基因产物的基本信息，这是长期研究其结构和功能所需的。重点是制备抗体和开发可靠的系统来表达全长或截短的蛋白质。具体目的是：(1)确定HfI-A27的组织分布，并在大肠杆菌或杆状病毒系统中表达，以用于抗体生产和结构研究。(2)在构成启动子或诱导型启动子的控制下，在猪或人细胞系中稳定表达该基因。(3)测定蛋白质大小、细胞定位和寡聚体结构。(4)启动研究以确定HfI-A27基因产物在HSV进入过程中的功能及其对人类细胞的天然作用(S)。长期目标是了解HSV感染是如何涉及多个细胞蛋白受体和病毒包膜蛋白的附着，从而在中性pH下导致膜融合。基于这些对HFI-A27的初步实验结果，并使用开发的细胞系和抗体，将在其他人类受体蛋白的背景下研究A27蛋白，以便于HSV进入及其自然的人类细胞功能(S)。实验结果结合对预测的蛋白质序列的计算机分析，应该可以形成假说，以推动对A27的进一步研究。这项研究对于了解病毒细胞进入时的相互作用以及确定一个新的人类基因及其未鉴定的蛋白质产物的自然功能具有重要意义。

DESCRIPTION (provided by applicant): This research initiates studies to characterize a newly isolated human gene currently designated human fetal lung cDNA A27 (hfI-A27). It is one of three novel human genes isolated in a functional screen of a human fetal lung cDNA library. We looked for genes that transfer susceptibility for HSV entry to porcine cells that lack a receptor for entry of HSV. The nucleotide sequence of hfl-A27 contains an open reading frame that produces in vitro translated proteins of 37kD and 28kD. Its gene product mediates HSV-1 entry when expressed transiently in porcine cells. Goals are to develop reagents and obtain fundamental information about the uncharacterized hfl- A27 gene product required for long term studies of its structure and functions. Priorities are preparation of antibody and development of reliable systems to express full length or truncated proteins. Specific aims are to: (1) Determine tissue distribution of mRNA and express hfI-A27 in an E.coli or baculovirus system for antibody production and studies of structure. (2) Express the gene stably in porcine or human cell lines under control of constitutive or inducible promoters. (3) Determine protein size, cellular location and oligomeric structure. (4) Initiate studies to determine function of the hfI-A27 gene product in HSV entry and its natural role(s) for human cells. The long-term objective is to understand how HSV infection involves multiple cellular protein receptors and viral envelope proteins for attachments that lead to membrane fusion at neutral pH. Based on results of these initial experiments with hfI-A27 and using cell lines and antibodies developed, the A27 protein will be studied in the context of other human receptor proteins for HSV entry and for its natural human cell function(s). Experimental results combined with computer analyses of the predicted protein sequence should allow formation of hypotheses to drive further studies of A27. The research is significant to understanding viral cell interactions at entry and to defining natural functions of one new human gene and its uncharacterized protein product.