 | Paul Reubens als Pee-wee Herman |
 | Penelope Ann Miller als Winnie |
 | Kris Kristofferson als Mace Montana |
 | Valeria Golino als Gina Piccolapupula |
 | Benicio del Toro als Duke the Dog-Faced Boy |
 | Wayne White als Vance the Pig (voice) |
 | Susan Tyrrell als Midge Montana |
 | Albert Henderson als Mr. Ryan |
 | Jack Murdock als Otis |
 | David Byrd als Deke |
 | Mary Jackson als Mrs. Dill |
 | Frances Bay als Mrs. Haynes |
 | Leo Gordon als Joe the Blacksmith (as Leo V. Gordon) |
 | Anne Seymour als Pearl |
 | Kenneth Tobey als Sheriff |
 | Stephanie Hodge als Mermaid |
 | Franco Columbu als Otto the Strongman |
 | Kevin Peter Hall als Big John |
 | Lynne Marie Stewart als Zelda the Bearded Lady |
 | Dustin Diamond als Child - Deke |
 | Jay Robinson als Cook |
| Kevin B. Kaplowitz als Child - Mr. Ryan |
| Jeffrey R. Shaw als Child - Otis |
| Marie Hawkins als Child - Pearl |
| Shea Joachim als Child - Sheriff |
| Molly Ann Carter als School Child |
| Myka Peck als School Child |
 | Patricia M. Peters als Web Girl |
 | Joey Arias als Shim |
 | Helen Siff |
 | Matthias Hues als Oscar - Lion Tamer |
| Vance Colvig als Clownie |
| April Tatro als Human Pretzel |
| Danny Elfman | Music |
| Paul Reubens | Producer |
| Paul Reubens | Writer |
| Jeff Gourson | Editor |
| George McGrath | Writer |
| Randal Kleiser | Director |
| Debra Hill | Producer |
| Steven Poster | Director of Photography |
| Robert Turturice | Costume Design |
| Victoria Thomas | Casting |
| Richard Edlund | Visual Effects Producer |
| Stephen Marsh | Production Design |
| William E. McEuen | Executive Producer |
| Richard Gilbert Abramson | Executive Producer |
| Bill Couch | Stunt Coordinator |
| Gary Morgan | Stunts |
| Karen Price | Stunts |
| Bill Couch Jr. | Stunts |
| Bob Yerkes | Stunts |
| Alex Plasschaert | Stunts |
| Kurt Neumann | Unit Production Manager |
| Roger Joseph Pugliese | First Assistant Director |
| Linda Montanti | Second Assistant Director |
| Beala Neel | Art Direction |
| Anne D. McCulley | Set Decoration |
| Stephen Homsy | Set Designer |
| Carl Aldana | Set Designer |
| Kenneth L. Westcott | Leadman |
| Oscar Delgadillo | Leadman |
| John Koester | Camera Operator |
| George Kohut | Camera Operator |
| Alan R. Disler | First Assistant Camera |
| Bing Sokolsky | First Assistant Camera |
| Bruce W. Talamon | Still Photographer |
| Mychal D. Smith | Boom Operator |
| Kirk Francis | Sound Mixer |
| James Plannette | Chief Lighting Technician |
| R. Michael De Chellis | Assistant Chief Lighting Technician |
| Jack Kirk | Assistant Chief Lighting Technician |
| Scott McKnight | Lighting Technician |
| Hank Sheppherd | Second Company Grip |
| Paul Threlkeld | Dolly Grip |
| Joseph Graham | Dolly Grip |
| Mike Schwake | Dolly Grip |
| Horst Grandt | Property Master |
| Oscar Flores | Assistant Property Master |
| Dianne Ryder-Rennolds | Additional Editing |
| Irvin Paik | Assistant Editor |
| Dick Bernstein | Music Editor |
| Cecelia Hall | Supervising Sound Editor |
| Ronald Jacobs | Supervising Sound Editor |
| R.J. Palmer | Sound Editor |
| Trevor Jolly | Sound Editor |
| Sandina Bailo-Lape | Sound Editor |
| Pamela Bentkowski | Foley Editor |
| Dessie Markovsky | Supervising ADR Editor |
| Judee Gustafson | Assistant Sound Editor |
| Donald O. Mitchell | Sound Re-Recording Mixer |
| Kevin O'Connell | Sound Re-Recording Mixer |
| Rick Kline | Sound Re-Recording Mixer |
| Dan Wallin | Scoring Mixer |
| William Ross | Conductor |
| Steve Bartek | Orchestrator |
| Joyce King | Script Supervisor |
| Marcy Bolotin | Unit Publicist |
| Patsy Swayze | Choreographer |
| Renee Milliken | Casting Assistant |
| Ve Neill | Makeup Artist |
| Edouard F. Henriques | Makeup Artist |
| Yolanda Toussieng | Hairstylist |
| Greg Cannom | Prosthetic Designer |
| Matt Sweeney | Special Effects Supervisor |
| George Stokes | Construction Coordinator |
| James F. Orendorff | Construction Foreman |
| Ray Rarick | Construction Foreman |
| Joel Marrow | Transportation Coordinator |
| Terry Hager | Color Timer |
