New human gene that mediates herpes simplex virus entry

描述（由申请人提供）：本研究开始研究新分离的人类基因，目前命名为人类胎儿肺cDNA A27 （hfI-A27）。它是在人类胎儿肺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进入敏感性转移到缺乏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阅读框，产生37kD和28kD的体外翻译蛋白。其基因产物在猪细胞中短暂表达时介导HSV-1进入。目标是开发试剂并获得未表征的hfl- A27基因产物的基本信息，以便对其结构和功能进行长期研究。重点是制备抗体和开发可靠的系统来表达全长或截断的蛋白质。具体目的是：(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，用于抗体产生和结构研究。(2)在组成型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)确定蛋白质大小、细胞位置和寡聚物结构。(4)开展研究，确定hfI-A27基因产物在HSV入侵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的附着物，这些附着物导致中性ph下的膜融合。基于hfI-A27的这些初始实验结果，并使用细胞系和开发的抗体，A27蛋白将在HSV进入的其他人类受体蛋白及其自然人类细胞功能的背景下进行研究。实验结果与预测的蛋白质序列的计算机分析相结合，应该允许形成假设，以推动对A27的进一步研究。该研究对于理解病毒细胞进入时的相互作用，以及确定一种新的人类基因及其未表征的蛋白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.