Hiram Chamberlain, Presbyterian minister, was born at Monkton, Vermont, on April 2, 1797. As a descendant of a New England family of Revolutionary War patriots, Protestant clergymen, physicians, and small farmers, he was raised in an intense religiosity unusual even for that era in the "Burned-Over District" of western Vermont and the New York frontier. In 1818 he made his profession of faith at Rev. Dr. Gardner Spring's Presbyterian church in New York City. Also that year he entered Middlebury College in Vermont, from which he graduated in 1822. Able to think of "but little except the duty of constant prayer and deep humiliation before God," he entered Andover Theological Seminary and dedicated himself to "missionary labors." After a year's study at Princeton, he graduated in 1825 from Andover, where he helped to establish the American Home Missionary Society. On October 16, 1825, he received ordination as a missionary from the New York Presbytery at Dr. Spring's church. For the next two decades (1825–45) he held numerous pastoral duties in Missouri, at churches in St. Louis, Dardonne, Boonville, New Franklin, Fayette, and St. Charles. He also served as an agent and promoter for Marion College, as well as editor and publisher of the St. Louis newspaper Herald of Religious Liberty.
The Presbyterian split in 1838 found Hiram decidedly in the old school. In addition, he severed his ties with the American Home Missionary Society the next year, apparently over the society's increasing support of the abolition movement. Though Chamberlain himself owned no slaves and seems to have disapproved of slavery, he, like many old-school ministers, believed that slaveholding was not forbidden in scripture and that the relation of master and slave was a civil and domestic institution in which the church had no power to legislate. He was also an ardent believer in the separation of church and state, dedicated "to wake up the attention of Protestants to the errors and evils of Roman Catholicism." Enlightenment, Chamberlain believed, came only through self-denial, noble resolution, and an unemotional adherence to Christian duty.
In 1845 he moved down the Mississippi River to Memphis, Tennessee, where he accepted new pastoral duties. Two years later he again moved sixty miles eastward to Somerville, where he agreed to oversee the building of a new church. Chamberlain moved to Brownsville, Texas, in 1850 to organize, build, and lead the first Protestant church on the lower Rio Grande. On February 23 he organized the First Presbyterian Church of Brownsville. With Melinda Rankin he opened an academy for young girls, Rio Grande Female Institute, in 1854. Chamberlain was an ardent secessionist and served as chaplain of the Third Texas Infantry throughout the Civil War. He accepted the surrender of the Confederacy as a sign of God's will that the Union remain intact.
On October 9, 1825, at Dorset, Vermont, he married Maria Morse, with whom he had three children, only one of whom, Henrietta King, lived to maturity. On April 19, 1836, at St. Charles, Missouri, Chamberlain married Sarah H. Wardlaw of Rockbridge County, Virginia; they had no children. At Pinckney, Missouri, on October 16, 1842, he married Anna Adelia Griswold of Wethersfield, Connecticut, who survived him and died at Brooklyn, New York, on November 24, 1882. They had seven children, five of whom lived to maturity. Chamberlain died on November 1, 1866, at the parsonage of his church in Brownsville.