Pocahontas' birth year is unknown, but some historians estimate it to have been around 1596. In A True Relation of Virginia (1608), Smith described the Pocahontas he met in the spring of 1608 as being "a child of ten years old". In a letter written in 1616, he again described her as she was in 1608, but this time as "a child of twelve or thirteen years of age".

Pocahontas is most famously linked to the English colonist Captain John Smith, who arrived in Virginia with a hundred other settlers in April 1607, at the behest of the London Company. After building a fort on a marshy peninsula poking out into the James River, the Englishmen had numerous encounters over the next several months with the people of Tsenacommacah, some of them friendly, some hostile. Then, in December 1607, while exploring on the Chickahominy River, Smith was captured by a hunting party led by Powhatan's younger brother (or close relative) Opechancanough and brought to Powhatan's capital at Werowocomoco. In his 1608 account, Smith describes a great feast followed by a long talk with Powhatan. He does not mention Pocahontas in relation to his capture, and claims that they first met some months later. Huber understands the meeting of Smith and Powhatan as the latter's attempt to bring Smith, and so the English, into his chiefdom: Powhatan offered Smith rule of the town of Capahosic, which was close to Powhatan's capital at Werowocomoco. The paramount chief thus hoped to keep Smith and his men "nearby and better under control".

Pocahontas's capture occurred in the context of the First Anglo-Powhatan War, a conflict between the Jamestown settlers and the Native Americans that began late in the summer of 1609. In the first years of war, the English took control of the James River, both at its mouth and at the falls. Captain Samuel Argall, in the meantime, pursued contacts with Native American groups in the northern portion of Powhatan's paramount chiefdom. The Patawomecks, who lived on the Potomac River, were not always loyal to Powhatan, and living with them was a young English interpreter named Henry Spelman. In March 1613, Argall learned that Pocahontas was visiting the Patawomeck village of Passapatanzy and living under the protection of the Weroance Iopassus (also known as Japazaws).

Current Mattaponi tradition holds that Pocahontas's first husband was Kocoum, brother of the Patawomeck weroance Japazaws, and that Kocoum was killed by the English after his wife's capture in 1613. Today's Patawomecks believe that Pocahontas and Kocoum had a daughter, Ka-Okee, who was raised by the Patawomecks after her father's death and her mother's abduction.

Pocahontas's feelings about Rolfe are unknown. They were married on April 5, 1614, by chaplain Richard Buck, probably at Jamestown. For two years they lived at Rolfe's plantation, Varina Farms, across the James River from Henricus. Their son, Thomas, was born on January 30, 1615.

Pocahontas and her husband, John Rolfe, had one child, Thomas Rolfe, who was born in January 1615. The following year, Thomas' parents travelled to London.

Although Pocahontas was not a Princess in the context of Powhatan culture, the Virginia Company nevertheless presented her as a Princess to the English public. The inscription on a 1616 engraving of Pocahontas, made for the company, reads: "MATOAKA ALS REBECCA FILIA POTENTISS : PRINC : POWHATANI IMP:VIRGINIÆ", which means: "Matoaka, alias Rebecca, daughter of the most powerful Prince of the Powhatan Empire of Virginia". Many English at this time recognized Powhatan to be the ruler of an empire, and they presumably accorded to his daughter what they considered appropriate status. Smith's letter to Queen Anne refers to "Powhatan their chief King". Cleric and travel Writer Samuel Purchas recalled meeting Pocahontas in London, noting that she impressed those she met because she "carried her selfe as the daughter of a king". When he met her again in London, Smith referred to Pocahontas deferentially as a "Kings daughter".

Pocahontas' funeral took place on March 21, 1617, in the parish of Saint George's, Gravesend. Her grave is thought to be underneath the church's chancel, though since that church was destroyed in a fire in 1727, her exact gravesite is unknown. Her memory is honored with a life-size bronze statue at St. George's Church by william Ordway Partridge.

In a later publication, True Travels (1630), Smith claimed a similar rescue by another young girl in 1602, following his capture by Turks in Hungary; the story resembles a popular contemporary type of moral tale, in which a Christian hero maintains his faith despite threats and intimidation. Karen Ordahl Kupperman suggests that Smith used such details to embroider his first account, thus producing a more dramatic, second account of his encounter with Pocahontas as a heroine worthy of reception by Queen Anne. Its later revision and publication was probably an attempt to raise his own stock and reputation; he had long since fallen from favor with the London Company, which had funded the Jamestown enterprise. Anthropologist Frederic W. Gleach, drawing on substantial ethnohistory, suggests that Smith's second account, while substantially accurate, represents his misunderstanding of a three-stage ritual intended to adopt Smith, as representative of the English colony, into the confederacy; but not all Writers are convinced, some suggesting the absence of certain corroborating evidence.

After her death, increasingly fanciful and romanticized representations of Pocahontas were produced, in which Pocahontas and Smith were romantically involved. Contemporary sources substantiate claims of their friendship, not romance. The first claim of their romantic involvement was in John Davis' Travels in the United States of America (1803)

Pocahontas has been considered in popular culture to be a Princess. In 1841, william Watson Waldron of Trinity College, Dublin, in Ireland, published Pocahontas, American Princess: and Other Poems, calling Pocahontas "the beloved and only surviving daughter of the king". Pocahontas was her father's "delight and darling", according to the colonist Captain Ralph Hamor but she was not in line to inherit a position as a weroance, subchief, or mamanatowick (paramount chief). Instead, Powhatan's brothers, sisters, and his sisters' children all stood in line to succeed him. In his A Map of Virginia John Smith explained how matrilineal inheritance worked among the Powhatans:

In 1907, Pocahontas became the first Native American to be honored on a US stamp. She was a member of the inaugural class of Virginia Women in History in 2000.

During the year-long wait, she was held at Henricus, in modern-day Chesterfield County, Virginia. Little is known about her life there, although colonist Ralph Hamor wrote that she received "extraordinary courteous usage". Linwood "Little Bear" Custalow, in a 2007 book, refers to an oral tradition that during this time, Pocahontas was raped; according to Helen Rountree, "Other historians have disputed that such oral tradition survived and instead argue that any mistreatment of Pocahontas would have gone against the interests of the English in their negotiations with Powhatan. A truce had been called, the Indians still far outnumbered the English, and the colonists feared retaliation."

The Virginia Company of London had long seen one of its primary goals as the conversion of Native Americans to Christianity. With the conversion of Pocahontas and her marriage to an Englishman – all of which helped bring an end to the First Anglo-Powhatan War – the company saw an opportunity to promote investment. The company decided to bring Pocahontas to England as a symbol of the tamed New World "savage" and the success of the Virginia colony. In 1616, the Rolfes travelled to England, arriving at the port of Plymouth on June 12. They journeyed to London by coach, accompanied by a group of about eleven other Powhatans, including a holy man named Tomocomo. John Smith was living in London at the time and while Pocahontas was in Plymouth, she learned he was still alive. Smith did not meet Pocahontas, but wrote to Queen Anne, the wife of King James, urging that Pocahontas be treated with respect as a royal visitor. He suggested that if she were treated badly, her "present love to us and Christianity might turn to ... scorn and fury", and England might lose the chance to "rightly have a Kingdom by her means".

According to the colonist william Strachey, "Pocahontas" was a childhood nickname that probably referred to her frolicsome nature; it meant "little wanton"; some interpret the meaning as "playful one". The 18th-century Historian william Stith claimed that "her real name, it seems, was originally Matoax, which the Indians carefully concealed from the English and changed it to Pocahontas, out of a superstitious fear, lest they, by the knowledge of her true name, should be enabled to do her some hurt." According to the Anthropologist Helen C. Rountree, Pocahontas "revealed [her secret name] to the English only after she had taken another religious—baptismal—name, Rebecca".

In July 2015, the Pamunkey Indian Tribe, descendants of the Powhatan chiefdom, of which Pocahontas was a member, became the first federally recognized tribe in the state of Virginia.