IIS 6.0 Default Document Not Working After .NET Framework 4.0 Install/Update
Google found us an article that identifies the problem in detail and explains how to apply a fix for it:
http://blogs.msdn.com/b/tmarq/archive/2010/06/18/how-to-disable-the-asp-net-v4-0-extensionless-url-feature-on-iis-6-0.aspx
Just add a DWORD registry key "HKEY_LOCAL_MACHINE\SOFTWARE\Microsoft\ASP.NET\4.0.30319.0\EnableExtensionlessUrls" with a value of "0" then restart IIS to fix the issue.