Comparative Studies of the Regeneration of HeLa Cell Receptors for Poliovirus T1 and Coxsackievirus B3

Abstract

Enterovirus receptors of live HeLa cells have been shown to reappear after inactivation by proteolytic enzymes, provided the cells are incubated at 37 C in a nutritionally adequate medium. Regeneration of receptor activity for poliovirus Ti occurred at a significantly faster rate than for coxsackievirus B3. The regenerative process for both types of receptors studied evidently required an active process of protein synthesis, since it was found that reappearance of receptor activity was inhibited by streptovitacin A, puromycin, and actinomycin D. Substitution of pfluorophenylalanine for the naturally occurring amino acid, at concentrations which inhibited virus synthesis, was without effect on regeneration of receptor activity. It is anticipated that these findings will aid in the study of the biosynthesis and subsequent chemical characterization of viral receptors of living host cells.

Virus-Receptor Interaction in an Adenovirus System

“…A clear difference is also shown by the fact that adenovirus fiber can block adenovirus uptake but does not affect poliovirus attachment (Table 5). In contrast to previous studies on virus-receptor interaction (17,25,32), neither the inhibition of virus infectivity nor hemagglutination-inhibition was sensitive enough to use as an assay system. It was possible to develop a sensitive and specific in vitro assay for adenovirus receptors based upon the stability of the virus-receptor complex in CsCl solutions in which the complex can be banded at about 1.2 g/cc.…”

“…The location of receptors at the plasma membrane has been established for picornavirus (17,32). Treatment of intact cells with trypsin and chymotrypsin, which destroy the receptors for polioand coxsackievirus (17,25), enhances the attachment rate for adenovirus (Fig. 1).…”

“…In other virus-cell systems the initial virusreceptor interaction appears to involve considerable specificity; for example, HeLa cell receptors for picornaviruses differ in susceptibility to enzymatic destruction and in the time required for their regeneration (17,25,32). The receptors for poliovirus appear also to be different from those of coxsackievirus, as revealed by cross-saturation experiments (17).…”

De‐sulfation of MG‐63 cell glycosaminoglycans delays in vitro osteogenesis, up‐regulates cholesterol synthesis and disrupts cell cycle and the actin cytoskeleton

“…Trypsinization of cells prior to seeding them enhanced the effects of chlorate, reducing cell proliferation by 60% compared to trypsinized cells that were not treated with chlorate. This is despite previous observations that membrane receptors can be regenerated within hours of trypsinization (Levitt and Crowell, 1967;McCune and Volsky, 1984;Borisuth et al, 1992). When the extracellular domains were left intact, we observed only a 30% drop in proliferation with chlorate treatment (Fig.…”

Properties of the deoxycholate-solubilized HeLa cell plasma membrane receptor for binding group B coxsackieviruses

“…The six serotypes of the group B coxsackieviruses bind to a common receptor on HeLa cells which is distinct from that binding the polioviruses, group A coxsackieviruses, and echoviruses (14). The receptors on intact HeLa cells are inactivated by chymotrypsin (22), and their regeneration requires host cell protein and mRNA synthesis (13). The development of a sodium deoxycholate (DOC) extraction procedure to solubilize biologically active, native receptors from HeLa cells (4,12) provided an impetus to further characterize them. The results presented herein represent the first characterization of soluble host cell receptors for binding group B coxsackieviruses.…”