We used monoclonal antibodies reacting with glycoproteins specified by herpes simplex virus type 2 (HSV-2) to characterize the individual antigens in terms of structure, processing, and kinetics of synthesis in BHK or Vero infected cells. Our results provided a direct demonstration of the structural identity of the gA and gB proteins of HSV-2 as well as confirmation of the existence of type-specific and type-common domains within the gD molecule. They also show that, with the exception of gC, processing of the viral glycoproteins differs to some extent in Vero and BHK infected cells, possibly as a result of different efficiency of glycosylation or different processing of underglycosylated and unglycosylated products in the two cell types. Finally, we showed that individual HSV-2 glycoproteins are synthesized at greatly different times during the infectious cycle, possibly in response to their different roles in virus replication and assembly.

Vitellins were identified, purified, and analyzed from insects representing eight orders. The structures and polypeptide constituents of vitellins of Hyalophora cecropia, Tenebrio molitor, Rhodnius prolixus, Forficula auricularia, Periplaneta americana, and a mayfly were found to have common features. The native proteins had Mr of 385,000-470.000 (385-470 K) and were composed of high (100-180 K) and low (47-84 K) molecular weight polypeptides in equimolarproportions. The vitellins of Apis mellifem, a sphecid wasp, and Aedes aegypti, however, had lower Mr (200-350 K) and were composed of only large polypeptides (170-190 K). The higher Diptera form a distinct third group with vitellins made up entirely of small polypeptides of about 50 K.Oogenesis in both insects and oviparous vertebrates is characterized by the accumulation of large quantities of yolk protein in the eggs. In insects, the female-specific yolk precursor proteins, vitellogenins, are synthesized in the fat body, secreted into the hemolymph, and se questered by the developing oocyte, sometimes after slight modifications, as vitellin (Telfer, '65; Wyatt and Pan, '78; Engelmann, '79; Hagedorn and Kunkel, '79). Vitellogenin synthesis in many insect species is under hormonal control, so that these systems can be used as models for the investigation of mechaqisms involved in hormonecontrolled gene expression (Hagedorn and Kunkel, '79; Engelmann, '74; Chen et al., '79).Several criteria have been used to define the vitellins. They are usually considered to be sexlimited proteins in that they are present in large quantities only in the female of a species (Pan et al., '69; Bell, '69); however, vitellogenin has been found in males of Rhodnius prolixus (Chalaye, '79) and Oncopeltus fasciatus (Kelly and Telfer, '77). In most species, vitellins comprise 60-90% of the total soluble egg yolk protein (Chenet al., '76; Hagedorn and Judson, '72; Bell, "70). Most hemolymph proteins can be found in the oocyte, but only vitellogenins are selectively concentrated by as much as 100-fold over their levels in the hemolymph (Pan, '71). Finally, the fat body is generally the principal, and perhaps exclusive, site of synthesis (Pan et al., '69), but in Drosophila species vitellogenins may also be synthesized in the ovary (Bownes and Hames, '78). Generally, vitellins are identified as sex-limited proteins that form a substantial proportion of the total soluble egg protein, and for only a few species have data been obtained for all the above four criteria.Although the physical and chemical proper- '77). but those of Drosophila species vary substantially from those of other insects (GeltiDouka et al., '74). The relationship of the constituent polypeptides to the native protein has been difficult to determine since simple stoichiometric relationships among the polypeptides have not previously been apparent. We have identified and characterized the vitellins of a number of insect species and report the existence of three definable groups. MATERIALS AND METHODS Insects

Monoclonal antibodies to herpes simplex virus type 2 were found to precipitate different numbers of radiolabeled polypeptides from lysates of virus-infected cells. Antibodies directed against two viral glycoproteins were characterized. Antibodies from hybridoma 17aA2 precipitated a 60,000-molecular-weight polypeptide which chased into a 66,000and a 79,000-molecular-weight polypeptide. All three polypeptides labeled in the presence of [3H]glucosamine and had similar tryptic digest maps. The 60,000-molecular-weight polypeptide also chased into a 31,000-molecular-weight species which did not label with [3H]glucosamine. Antibodies from hybridoma 17,8C2 precipitated a 50,000-molecular-weight polypeptide which chased into a 56,000and an 80,000-molecular-weight polypeptide. These polypeptides also shared a similar tryptic digest map and labeled with [3H]glucosamine. Both monoclonal antibodies were herpes simplex virus type 2 specific. The viral proteins precipitated by 17aA2 antibodies had characteristics similar to those reported for glycoprotein E, whereas the proteins precipitated by 17,BC2 antibodies appeared to represent a glycoprotein not previously described. This glycoprotein should be tentatively designated glycoprotein F.

No abstract

Primer extension of Pichinde arenavirus purified virion RNA suggests that genomes have at least a single nontemplated base at the 5' end which is a G in all cDNA clones having one such single base. On the other hand, the predominant products of primer extension on total virus-infected-cell RNA are at positions -1 and -2. The primer extension product at position -2 is not represented in virion RNA, and neither of these products is proportionally represented in mRNA. mRNA is predominantly 3 or 4 bases longer than genomes and antigenomes, but primer extension products as long as 7 bases were observed. The sequence of nontemplated bases reported here is unambiguous with respect to the 5'-terminal base and supports the view that there is a sequence preference for a G at the 5' termini of mRNAs. Assessment of our sequence data in the context of the sequences of Tacaribe and lymphocytic choriomeningitis viruses suggests that the mechanism of initiation of arenavirus transcription is fundamentally different from that of members of the families Orthomyxoviridae and Bunyaviridae.