New human gene that mediates herpes simplex virus entry

描述（由申请人提供）：本研究启动了对一种新分离的人类基因（目前命名为人胎肺cDNA A27（hfI-A27））进行表征的研究。 它是在人胎肺cDNA文库的功能筛选中分离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的核苷酸序列包含一个开放的阅读框架，其在体外产生37 kD和28 kD的翻译蛋白。 其基因产物在猪细胞中瞬时表达时介导HSV-1进入。 目的是开发试剂并获得长期研究其结构和功能所需的未表征的hf 1- A27基因产物的基本信息。优先考虑的是抗体的制备和可靠的系统表达全长或截短的蛋白质的发展。 具体目标是：（1）确定mRNA的组织分布，并在大肠杆菌或杆状病毒系统中表达hfI-A27，用于抗体生产和结构研究。 (2)在组成型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)确定蛋白质大小、细胞位置和寡聚体结构。 (4)启动研究以确定hfI-A27基因产物在HSV进入中的功能及其对人类细胞的天然作用。 长期目标是了解HSV感染如何涉及多种细胞蛋白受体和病毒包膜蛋白，用于在中性pH下导致膜融合的附着。基于hfI-A27的这些初始实验结果，并使用开发的细胞系和抗体，将在HSV进入的其他人类受体蛋白及其天然人类细胞功能的背景下研究A27蛋白。 结合计算机分析预测的蛋白质序列的实验结果应该允许形成的假设，以推动A27的进一步研究。 这项研究对于理解病毒进入细胞时的相互作用以及确定一个新的人类基因及其未表征的蛋白质产物的天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.