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- Captain Augustus "Gus" McCrae — Co-owner of the Hat Creek Cattle Company and a Captain of the Texas Rangers, McCrae considers himself the brains of the outfit. Generous, humorous, and lazy to the point of eccentricity, he serves as a foil to the more serious, practical Call. When not working (which he does as little as possible), Gus pursues his three chief interests in life: women, alcohol, and cards. He is well known in the territory for his loud voice, superior eyesight, and accuracy with a revolver.
- Captain Woodrow F. Call — Gus's partner in the company. Less verbose and chatty than McCrae, Call works long and hard and sees no reason why others should not do the same. A former Texas Ranger who won a merit award from the Governor of Texas for "courage under fire", he served with Gus when both were young men. Though Call has utter disdain for lazy men who drink, gamble, and whore their lives away, he has his own failings, in particular his unwillingness to admit he is Newt's father. Call's ability to "break" unmanageable horses is also well known.
- Pea Eye Parker — The wrangler and blacksmith of the Hat Creek Cattle Company, Pea Eye served as a corporal in the Rangers under Gus and Call. Pea Eye (his real name long forgotten) is not especially bright, but he is reliable, brave, and kind. He follows Call's lead without question.
- Joshua Deets — An ex-slave and former Ranger, Deets is a ranch hand at the company. On the drive, he serves as scout. A remarkable tracker and morally upright man, he is one of the few men whom Call respects and trusts.
- Newt Dobbs — An orphan raised by Gus and Call. His mother was a prostitute named Maggie Tilton, who died when he was a child. He knows his mother was a prostitute, but has no idea who his father might be. Most observers, notably Gus and Clara Allen, are quite certain that Call is his father. Call apparently knows this too, but is never able to publicly admit it. Call does give Newt his horse "The Hell Bitch" which he values far more than his name. As McMurtry remarks in the interview accompanying the DVD, Newt is the "Lonesome Dove" from which the title comes.
- Jake Spoon — A former comrade-in-arms of Gus, Call, Pea Eye, and Deets. Jake is, if anything, lazier than Gus, but without the latter's redeeming virtues. A gambler and drinker, Jake prefers living in luxury and ease and shirks work with a passion, which irks Call mightily. He is, however, a man of great personal charm and is seldom unlucky in love.
- Dishwater "Dish" Boggett — A cowboy of great skill, "Dish" is the top hand for Call's cattle drive. His main aspiration is to win the love of Lorena Wood. His nickname derives from having swallowed dishwater, being so thirsty that he could not wait for the water barrel.
- Lorena Wood Parker — A kind-hearted young woman who was forced into prostitution by her lover, she was then abandoned in Lonesome Dove. Lorena is taciturn, strong-willed, and intimidating, refusing to submit meekly to her various admirers. Discontent with her line of work, "Lorie" hopes to leave the dead town and find her way to San Francisco.
- Blue Duck — When Gus and Call quit Rangering, Blue Duck was unfinished business. The son of a Comanche war chief and his Mexican prisoner, Blue Duck leads a band of renegade Indians and buffalo hunters. He is feared across the plains as a murderer, rapist, and slaver.
- July Johnson — The sheriff of the town of Fort Smith, Arkansas. July is a kind, long-suffering young man, recently married to a woman he knows little about and who is utterly disdainful of his attention. After his brother, Ben, is accidentally killed by Jake Spoon, July's domineering sister-in-law Peach bullies him into setting out in pursuit. July is accompanied by his young stepson, Joe, and his incompetent deputy, Roscoe. (In the 1968 film Bandolero!, the sheriff, played by George Kennedy, is named July Johnson and his deputy, played by Andrew Prine, is named Roscoe.)
- Roscoe Brown — The deputy sheriff of Fort Smith, Arkansas. Roscoe is a timid man who spends his days playing dominoes and occasionally bringing in the local drunk for an overnight stay at the jail. After July is sent off after Jake Spoon, Roscoe is coerced by July's sister-in-law, Peach, into tracking down July who left to track down Jake Spoon who killed Peach's husband in Ft. Smith. Roscoe is also afraid of wild pigs. He, Joe, and a girl named Janey are later killed by Blue Duck. Roscoe's sidearm is a Colt Patterson 1836.
- Clara Allen — A former love of Gus, she declined his marriage proposals years ago. She lives in Nebraska, married to a Bob Allen, a horse trader who is comatose, having been kicked in the head by a horse. They have two girls, though she is afflicted deeply by the death of her sons. Though separated from Gus by many miles and years, she still holds him fondly in her heart. In contrast, she has utter contempt for Call.
- Po Campo — Cook for the Hat Creek Cattle Company on their cattle drive. Picked up on the way during a stop in Austin (San Antonio in the miniseries). He is most notable for his use of "exotic" ingredients and his refusal to ride animals. Po had a wife that, in his words, "lives in Hell, where I sent her." His sons were previously killed by the renegade half-breed outlaw, Blue Duck.
- Elmira Johnson — July's coldhearted, pregnant wife. Shortly after July departs to track Jake Spoon, Elmira flees town in search of old flame Dee Boot. She finally gets to Ogallala just before Dee is hanged for murder. Along the way she travels on a whiskey boat, enlists a couple of buffalo hunters in her quest, and gives birth at Clara Allen's ranch then abandons her baby there. She and one of the buffalo hunters, Big Zwey, are killed by the Sioux shortly after leaving Ogallala.
- Peach Johnson — July Johnson's sister-in-law. She pressures July to bring Jake Spoon back for killing her husband. A week after July and Joe leave, she pressures Roscoe to find July and tell him his wife has run off and is pregnant.
- Bolivar — The cook for the Hat Creek Cattle Company. A former Mexican bandit / rustler who, in his retirement years, became a cook, a more relaxing, less dangerous occupation. (Nevertheless, his ability to provide goat meat for dinner — an animal that the Hat Creek Cattle Company does not raise — seems to point to the fact that he keeps himself in shape by still practicing some rustling on the side). He is obsessed with loudly and unnecessarily ringing the dinner bell to call the company for dinner, much to Gus's annoyance.
- Hugh Auld — Former trapper and friend of the Blood Indians who cause Augustus' death; finds Augustus as he's trying to reach Miles City and lends his horse so that Augustus can make it. Later joins the Hat Creek outift as they winter on the ranch.
- Jimmy and Ben Rainey, Bill and Pete Spettle, Soupy Jones, Needle Nelson, Jasper Fant, Bert Borum, Lippy Jones, Sean and Allen O'Brien — Other hands hired by Call to work the cattle drive.