Starting in 1959, he found steady work at Berry Gordy's Hitsville U.S.A. studio, home of the Motown record label.

[6][7][8] He played bass on the Smokey Robinson single "Way Over There" (1959), John Lee Hooker album Burnin' (1962) and the Reflections' "(Just Like) Romeo and Juliet" (1964).

For many years, they maintained a schedule of recording during the day at Motown's small basement "Studio A" (which they nicknamed "the Snakepit"), then playing gigs in jazz clubs at night.

For most of their career, however, the Funk Brothers went uncredited on Motown singles and albums, and their pay was considerably less than that received by the main artists or the label, hence their occasional freelance work elsewhere.

Eventually, Jamerson was put on retainer for $1,000 a week (US$8,762 in 2023 dollars[13]), which afforded him and his expanding family a comfortable lifestyle.

[14][15] His work includes hits such as, among hundreds of others, "You Can't Hurry Love" by the Supremes, "My Girl" by the Temptations, "Shotgun" by Jr. Walker & the All Stars, "For Once in My Life" and "I Was Made to Love Her" by Stevie Wonder, "Going to a Go-Go" by the Miracles, "Dancing in the Street" by Martha and the Vandellas, "I Heard It Through the Grapevine" by Gladys Knight & the Pips and later by Marvin Gaye, and most of the album What's Going On by Marvin Gaye, "Reach Out I'll Be There" and "Bernadette" by the Four Tops.

[19][20][c] According to fellow Funk Brothers in the 2002 documentary Standing in the Shadows of Motown, Gaye was desperate to have Jamerson play on "What's Going On", and went to several bars to find the bassist.

[23][24][d] He performed on 23 number-one hits on the pop charts,[e] a record narrowly surpassed only by Paul McCartney of the Beatles, who cites Jamerson as his biggest influence.

By contrast, many of Jamerson's basslines relied heavily on chromatic runs, syncopation, ghost notes and inversions, with frequent use of open strings.

[33] Jamerson's background as a jazz musician and upright bassist informed his playing style, and over time his technique and improvisational approach became more nuanced.

[1][34][35] Early examples of Jamerson's impact are "Rescue Me" by Fontella Bass and "You Won't See Me" and "Nowhere Man" by the Beatles.

[36](pp144–146) In a 1983 interview with Musician magazine, Jamerson said Motown's songwriting-production team "would give me the chord sheet, but they couldn't write for me.

"[17][g][h] Bassists who have noted Jamerson's contribution or been influenced by him include Rocco Prestia, Anthony Jackson, Pino Palladino, Paul McCartney, Bob Babbitt, Nathan Watts, Will Lee, Geddy Lee, Chuck Rainey, Marcus Miller, Mike Mills, Phil Chen, John Entwistle, Michael League, Mike Watt, Sting, John Paul Jones, Bernard Odum, Victor Wooten, Robert DeLeo, Glenn Hughes, Tommy Shannon, Suzi Quatro, Ron Asheton, Tony Sales, Peter Cetera, Robert Kool Bell, Bootsy Collins, Flea, Jaco Pastorius, Stanley Clarke, Michael Henderson, Jack Bruce, John Patitucci, Jason Newsted, Rick Danko, Garry Tallent, Alan Gorrie, Jerry Jemmott, Andy Fraser, Brian Wilson and others.

[16] He also played on recordings by Robert Palmer (Pressure Drop, 1975), Dennis Coffey (Instant Coffey, 1974), Wah Wah Watson (Elementary, 1976), Rhythm Heritage (1976), Al Wilson (1977),[7] Dennis Wilson (Pacific Ocean Blue, 1977), Eloise Laws (1977), Smokey Robinson (1978), Ben E. King (1978),[40] Hubert Laws (1979), Tavares (1980), Joe Sample/David T. Walker (Swing Street Cafe, 1981), and Bloodstone (1982).

[23][17] As other musicians went on to use high-tech amplifiers, round-wound strings, and simpler, more repetitive bass lines incorporating new techniques like slapping, Jamerson's style fell out of favor with local producers as he was reluctant to try new things.

[6](p49) Long troubled by alcoholism, Jamerson died of complications from cirrhosis, heart failure and pneumonia on August 2, 1983, in Los Angeles.

[49] He received a Grammy Lifetime Achievement Award in 2004,[50] and he was inducted into the Musicians Hall of Fame in 2007 both as a member of the Funk Brothers.

[62] Also, the South Carolina Senate, the House of Representatives and the town of Edisto Island have passed resolutions in recognition of his contributions.

After graduating from high school, he bought a German upright bass which he later used on such Motown hits as "My Guy" by Mary Wells and "Heat Wave" by Martha and the Vandellas.

It had a three-tone sunburst finish, a tortoiseshell pickguard, rosewood fretboard and chrome pickup and bridge covers (the latter containing a piece of foam used to dampen sustain and some overtones).

Another aspect of Jamerson's upright playing that carried over was his use of open strings, a technique long used by jazz bass players, to pivot around the fretboard which served to give his lines a fluid feeling.

[53] Jamerson's amplifier of choice at club performances was an Ampeg B-15; in larger venues, he used a blue Kustom with twin 15-inch speakers.

[32] On most of his studio recordings, his bass was plugged directly into the custom-made mixing console together with the guitars from Eddie Willis, Robert White and Joe Messina.