It takes more than just breaking and entering for a virus to successfully invade a cell. Getting to the cells centerwhere the host cells machinery will be co-opted to make more virusrequires navigating obstacles such as membranes and avoiding being recognized and kicked out by the host.
Now scientists from the
The research appears in the Dec. 28 issue of the journal Molecular Cell.
Led by Daniel Hebert, the UMass Amherst team was investigating Simian Virus 40 (SV40), a small, golf-ball shaped virus from the genus known as polyomavirus. Polyomaviruses are nonenveloped virusesthey lack the protective envelope that surrounds some virusesand SV40 has become the workhorse of scientists trying to understand how nonenveloped DNA viruses work.
Hebert began investigating the SV40 virus to better understand the quality control mechanisms of cells that the virus infects. Comprising only 6 proteins and some DNA, SV40 is what Hebert describes as a dinky little virus. Yet, he says, They know more about the cell than we dothey are surfing down the cellular pathways that we are trying to understand.
Previous research had detailed part of the virus journey to a cells interiorSV40 made it into the host cells endoplasmic reticulum (ER), a maze-like network of membranes where, among other things, proteins, carbohydrates and fats are put together. But how the virus genome got from the ER to the nucleusa necessary feat because viruses cant copy their own genetic material and must use the machinery of their hostwasnt clear.
To investigate, researchers had tried deleting the genes that encoded SV40s proteins. Of particular interest were the structural proteins known as VP1, VP2 and VP3. Looking at a mature SV40 virus from the outside, one would only see repeated bits of VP1, which make up the exterior of the golf-ball shaped virus. VP2 and VP3 are bound to the interior part of VP1, but on the insideone could only find them by taking the virus apart. Studies that deleted the genes for these viral proteins had conflicting resultssometimes SV40 was still infectious, in other cases it was not.
So Hebert and his graduate student Robert Daniels decided to couple experiments that knocked out the genes for the viral proteins with experiments that let them follow what exactly happened to virus once inside the host cells ER. First his team created mutant SV40svirus that couldnt make VP2, virus that couldnt make VP3 and virus that couldnt make VP2 or VP3. None of the mutants were capable of infecting cells.
The research team also took a closer look at each of the viral proteins and how they operated, both on their own and as a unit. Generally, proteins are either of the membrane persuasion or they are notthose with the chemical and physical properties that allow them to insert into a membrane arent usually found outside of membranes, and proteins with properties that make them adverse to membranes stay out of them. But Hebert and Daniels discovered a strange thing. When VP2 and VP3 were attached to VP1, as they are in the mature virus, none are membrane proteins. But when VP2 and VP3 are on their own, they can slice into membranes, potentially making a pore through which viral DNA could travel to the nucleus.
A protein that can make a hole in a membrane can be a self-defeating weapon for a virus, says Hebert. Slicing membranes before the hosts machinery makes lots more virus means death for the host cell without much viral progeny. But SV40 unleashes V2 and V3 with exquisite timing. The proteins are hidden inside the virus unable to make any holes until they get to the host cells ER.
The virus has perfect timing on the construction end as wellonce the host starts making more virus, the genes that code for VP1 get turned on firstthus when VP2 and VP3 are made, they can be snatched up by VPI before they can poke themselves through any membranes.
All mature polyomavirus have VP2 and VP3 tucked into their core, suggesting that all polyomavirus use the same secret weapon to gain access to the nucleus, says Hebert. While SV40 doesnt normally infect humans (it has been found in humans and was likely introduced in the 1950s via a contaminated polio vaccine), its compatriots, the polyomaviruses known as JC virus and BK virus do, and sometimes fatally. These new findings may lead to new strategies for treating or preventing infection by polyomavirus.
Hebert, whose background is in cellular biology, intends to keep studying SV40 to better understand the inner workings of the cells the virus infects. Viruses provide a valuable window to the cell to follow fundamental cellular processes and we can learn a lot by uncovering how these pathogens exploit their cellular host.
Source: University of Massachusetts, Amherst