Dennis Milton Chambers was born on May 9, 1959, in Baltimore, Maryland. He started on the drums at four and was working Baltimore-area nightclubs by six, after the bassist of a local jazz band called The Fingertips heard him playing while stuck in traffic. His mother, a former Motown backing singer who had formed a band after moving back to Baltimore, rehearsed in the family's apartment, and watching the band's drummer was, he has said, the one thing that kept him still as a child. He never studied formally and does not read music. At 13 he was offered a place in James Brown's band; his mother declined because Brown would not pay for a tutor on the road, and Chambers has said he is glad she did.

In 1978, at 18, he joined George Clinton's Parliament-Funkadelic organization by way of the Brides of Funkenstein, and he stayed with the P-Funk family until 1985. Discogs credits him as one of several drummers on Parliament's Gloryhallastoopid (1979), Funkadelic's Uncle Jam Wants You (1979), the Brides of Funkenstein's Never Buy Texas from a Cowboy (1979) and Clinton's Some of My Best Jokes Are Friends (1985). From 1981 he was also a house drummer for the Sugar Hill label and played on many of its records, although, as he told Drumeo in 2017, he is not the drummer on "Rapper's Delight," despite the long-running belief that he is.

The move into jazz-fusion came in 1986, when he joined the band of guitarist John Scofield. Scofield had seen him with the group Special EFX and asked him in for a record date; Chambers has said he did not realize at first that he was joining the band. Blue Matter (recorded September 1986, released 1987), Pick Hits Live (recorded in Tokyo on October 7, 1987) and Loud Jazz (recorded December 1987) are the three Scofield albums with Chambers and bassist Gary Grainger. From 1989 to 1992 he was in a touring group led by guitarist Mike Stern with saxophonist Bob Berg and bassist Lincoln Goines, heard on Odds or Evens (1991), and in 1992 he played on Return of the Brecker Brothers (GRP), the reunion of Randy Brecker and Michael Brecker after a decade apart. His own first records as a leader, the Big City EP (1991) and Getting Even (1992), appeared in the same years, followed by Outbreak (2002), Planet Earth (2005) and Groove and More (2013).

The 1990s widened the circle. In December 1993 he recorded Tokyo Live at the Blue Note in Tokyo with John McLaughlin and organist Joey DeFrancesco as the Free Spirits trio, and he went on to play on McLaughlin's The Promise (1995), The Heart of Things (1997) and Industrial Zen (2006). He toured with Steely Dan in 1993 and 1994 and is heard on the live album Alive in America (1995). In 1996 he formed the organ trio Niacin with bassist Billy Sheehan and keyboardist John Novello, which made a run of albums that included High Bias (1998) and Krush (2013). He cut the trio album Front Page (2000) with guitarist Biréli Lagrène and bassist Dominique Di Piazza, and in 2000 he became one of the three founders of CAB with bassist Bunny Brunel and guitarist Tony MacAlpine; CAB 2 (2001) was nominated for Best Contemporary Jazz Album at the 44th Annual Grammy Awards, his only Grammy nomination. Other projects of the period include Extraction (2003) with guitarist Greg Howe and bassist Victor Wooten, Boston T Party (2006) with bassist Jeff Berlin, guitarist David Fiuczynski and keyboardist T Lavitz, and the trio HBC with Scott Henderson and Berlin (2012). He also toured with Carlos Santana for more than a decade. Paris/New York, a trio album with pianist Thierry Maillard and bassist John Patitucci, was Maillard's 1998 debut, released in Europe at the time and issued in the United States by Ghost Note Records in 2022.

Modern Drummer inducted him into its Hall of Fame in 2001, Berklee College of Music gave him an honorary doctorate of music in December 2004, and in 2016 Rolling Stone placed him at number 63 on its list of the 100 greatest drummers of all time. In July 2014 he was taken ill during a tour, a week after playing the Festival da Jazz in St. Moritz with Stern, and spent time in intensive care; by his own account he did not touch a drum kit for a year, and his first gig back was with Stern at Blues Alley in Washington. He has remained active since: he was with Scofield's band at the Kongsberg Jazz Festival in Norway in 2017, is the drummer on three tracks of Stern's Echoes and Other Songs (2024), plays on Oz Noy's Triple Play (2023), and toured with Stern in Australia in February 2025 and on the US West Coast in December 2025.

Chambers is a groove drummer with a fusion drummer's technique: a deep, backbeat-heavy pocket learned in P-Funk, joined to speed, independence and a taste for metric displacement that made him a first call for fusion bandleaders. AllMusic describes his style as propulsive and versatile enough for both small combos and large groups, and German Wikipedia calls him a world-famous jazz and fusion drummer known for an exact, powerful technique. He is self-taught, and he has said he plays without written parts and sometimes hears a Coltrane, Holdsworth or McLaughlin melody in his head during a solo and lets it displace the beat, which is why bassists who try to follow him through a solo can lose their place. He names Clyde Stubblefield, Al Jackson Jr., Steve Gadd, Vinnie Colaiuta, Gary Husband, Jack DeJohnette, Billy Cobham, Buddy Rich, Elvin Jones, Roy Haynes and Tony Williams among his influences. His long run with guitarists such as John Scofield, Mike Stern and John McLaughlin rests on the same trait: he supplies a steady, funky floor and leaves the guitarist free to move. He plays Pearl drums and Zildjian cymbals and has endorsed both brands for years. Beyond the Modern Drummer Hall of Fame and the Rolling Stone ranking, his standing among drummers shows in the number of peers, including Billy Hart and Billy Cobham, who turn up at his gigs. His history includes P-Funk, Santana and Steely Dan, so he is often described as a crossover figure, but the great majority of his recorded work is jazz-fusion and jazz-funk, and English Wikipedia files him under American jazz drummers.